CCL4 (MIP-1-beta, ACT-2, LAG-1) is a small secreted CC chemokine produced by activated T lymphocytes, monocytes/macrophages, NK cells, dendritic cells and other leukocytes. It is synthesized as a 92-residue precursor with a signal peptide; after secretion the N-terminal two residues are frequently removed to give MIP-1-beta(3-69), which is the predominant natural form. The mature protein adopts the canonical chemokine fold, forms homodimers and heterodimers with CCL3/MIP-1-alpha, and reversibly polymerizes into rod-shaped double-helical polymers that bury the receptor-binding surface and protect the chemokine from proteolysis, thereby shaping the chemotactic gradient. Its principal receptor is CCR5, a Gi-coupled GPCR: ligand binding triggers calcium mobilization, cytoskeletal polarization and directed migration of CCR5-bearing cells, chiefly monocytes/macrophages, effector and memory T cells, NK cells and immature dendritic cells, and also drives rapid internalization and down-modulation of the receptor. The naturally truncated MIP-1-beta(3-69) broadens receptor usage to CCR1 and CCR2 isoform B. CCL4 is a central mediator of the inflammatory response, recruiting leukocytes to sites of infection and tissue damage and promoting leukocyte adhesion and NK-target cell interactions. Because it occupies and down-regulates CCR5, CCL4 is also one of the three major HIV-suppressive factors released by CD8+ T cells and restricts entry of R5-tropic HIV-1 into target cells. Reported ligand activity at CCR8 was not reproduced and is not considered established.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005125 cytokine activity | TAS PMID:2462251 Identification, cloning, and characterization of an immune a... | ACCEPT | Summary: Original Act-2 cloning paper showing that this activation-induced gene encodes a secreted product proposed to be a cytokine. CCL4 is a chemokine, a subclass of cytokines. Reason: Cytokine activity is a correct, if general, parent of chemokine activity (GO:0008009), which is also annotated and is the core term. Supporting Evidence: PMID:2462251 It is therefore possible that Act-2 represents a newly discovered cytokine. file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. |
| GO:0005515 protein binding | IPI PMID:25416956 A proteome-scale map of the human interactome network. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC30A2 (Q9BRI3), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC30A2 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:25416956 Here, we describe a systematic map of |
| GO:0005515 protein binding | IPI PMID:31515488 Extensive disruption of protein interactions by genetic vari... | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC30A2 (Q9BRI3), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC30A2 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:31515488 we then performed yeast two-hybrid (Y2H) experiments to measure the impact of these 2009 missense SNVs across 2185 human proteinβprotein interactions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC7A1 (P30825), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC7A1 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and CNR2 (P34972), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: CNR2 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and GJA8 (P48165), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: GJA8 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC18A1 (P54219-3), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC18A1 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC10A6 (Q3KNW5), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC10A6 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and ARL13B (Q3SXY8), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: ARL13B is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and MRM1 (Q6IN84), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: MRM1 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and TMEM179B (Q7Z7N9), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: TMEM179B is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and UNC93A (Q86WB7-2), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: UNC93A is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC30A8 (Q8IWU4), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC30A8 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and TMEM86B (Q8N661), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: TMEM86B is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and GPR152 (Q8TDT2), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: GPR152 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and SLC30A2 (Q9BRI3), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: SLC30A2 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and TIMMDC1 (Q9NPL8), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: TIMMDC1 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and TMEM14B (Q9NUH8), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: TMEM14B is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and TAS2R5 (Q9NYW4), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: TAS2R5 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and GPRC5D (Q9NZD1), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: GPRC5D is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005515 protein binding | IPI PMID:32296183 A reference map of the human binary protein interactome. | REMOVE | Summary: Binary yeast two-hybrid interaction between CCL4 and MARCHF2 (Q9P0N8), reported by a CCSB proteome-scale interactome screen. No follow-up study links the two proteins functionally. Reason: Generic protein binding carries no functional information, so the annotation is uninformative rather than wrong; removal is not a claim that the reported interaction is false. No more specific molecular function can be derived from a screen hit: MARCHF2 is an intracellular or integral-membrane protein with no known role in chemokine biology, whereas CCL4 is a secreted CCR5 agonist, so there is no biological context for the pair. CCL4's informative binding activities are already captured by GO:0031730 CCR5 chemokine receptor binding and GO:0048020 CCR chemokine receptor binding. Supporting Evidence: PMID:32296183 We screened this search space a total of nine times with a panel of three Y2H assay versions |
| GO:0005576 extracellular region | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetic inference of extracellular localization across the CC chemokine family. CCL4 has a signal peptide and is secreted. Reason: CCL4 is a secreted chemokine acting on cell-surface receptors of target leukocytes; extracellular region is correct. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | IEA GO_REF:0000120 | ACCEPT | Summary: Electronic mapping of extracellular localization from InterPro chemokine signatures and the UniProt Secreted subcellular location. Reason: Correct; CCL4 is secreted. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | TAS PMID:2521882 Mitogenic activation of human T cells induces two closely re... | ACCEPT | Summary: The pAT 744 (CCL4) cloning paper describes a hydrophobic N-terminal leader characteristic of secreted proteins. Reason: Correct; CCL4 is secreted. Supporting Evidence: PMID:2521882 The predicted peptides encoded by these two clones feature hydrophobic N-terminal leaders characteristic of secreted proteins. |
| GO:0005576 extracellular region | TAS Reactome:R-HSA-380073 | ACCEPT | Summary: Reactome places extracellular CCL4 as a ligand of a Gi-activating chemokine GPCR. Reason: Correct; CCL4 is secreted and acts extracellularly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | TAS Reactome:R-HSA-443986 | ACCEPT | Summary: Reactome places extracellular CCL4 among inflammatory CC chemokines bound by the scavenger receptor ACKR2. Reason: Correct; CCL4 is secreted and acts extracellularly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | TAS Reactome:R-HSA-6784160 | ACCEPT | Summary: Reactome lists CCL4 among extracellular inflammatory mediators negatively regulated by IL10. Reason: Correct; CCL4 is secreted and acts extracellularly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | TAS Reactome:R-HSA-749454 | ACCEPT | Summary: Reactome places extracellular CCL4 in a ligand-GPCR-Gi complex. Reason: Correct; CCL4 is secreted and acts extracellularly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0005576 extracellular region | TAS Reactome:R-HSA-749456 | ACCEPT | Summary: Reactome places extracellular CCL4 as the ligand of a Gi-activating GPCR. Reason: Correct; CCL4 is secreted and acts extracellularly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SUBCELLULAR LOCATION: Secreted. |
| GO:0006954 inflammatory response | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetically inferred involvement of CC chemokines in the inflammatory response. Reason: CCL4 (MIP-1beta) is a pro-inflammatory chemokine produced by activated monocytes and lymphocytes that recruits CCR5-expressing leukocytes. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. |
| GO:0006954 inflammatory response | TAS PMID:2521882 Mitogenic activation of human T cells induces two closely re... | ACCEPT | Summary: Early cloning paper placing pAT 744 (CCL4) in a family of secreted factors associated with inflammatory responses. Reason: Consistent with the established inflammatory role of CCL4; duplicates the IBA annotation. Supporting Evidence: PMID:2521882 Some of these factors have been shown to display functions associated with an inflammatory response |
| GO:0006955 immune response | IEA GO_REF:0000002 | ACCEPT | Summary: InterPro2GO mapping from chemokine domain signatures. Reason: Broad but correct; more specific terms are also annotated. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. |
| GO:0006955 immune response | TAS PMID:10929056 Induction of interactions between CD44 and hyaluronic acid b... | ACCEPT | Summary: MIP-1beta is among pro-inflammatory mediators that rapidly activate CD44 on human T cells. Reason: Broad but correct. Supporting Evidence: PMID:10929056 chemokines (e.g. MIP-1beta, interleukin-8, and RANTES) |
| GO:0007155 cell adhesion | TAS PMID:10929056 Induction of interactions between CD44 and hyaluronic acid b... | MODIFY | Summary: A 2-3 h exposure of human T cells to MIP-1beta induced CD44-dependent adhesion of those T cells to immobilized hyaluronan under static and shear-stress conditions. Reason: CCL4 is not itself an adhesion molecule. It acts upstream, through chemokine receptor signaling on the T cell, to switch CD44 into a hyaluronan-binding state; hyaluronan is an extracellular matrix component, so the accurate term is positive regulation of cell-matrix adhesion. This is a downstream consequence of chemokine signaling rather than a core function. Proposed replacements: positive regulation of cell-matrix adhesion Supporting Evidence: PMID:10929056 is regulated by the cross-linking of CD3, cytokines (e.g. interleukin-2 and tumour necrosis factor-alpha), and chemokines (e.g. MIP-1beta, interleukin-8, and RANTES) |
| GO:0007163 establishment or maintenance of cell polarity | TAS PMID:9759849 Roles of chemokines and receptor polarization in NK-target c... | KEEP AS NON CORE | Summary: Chemokines produced in NK-target cell conjugates drive NK-cell polarization, with CCR2 and CCR5 clustering at the leading edge and adhesion molecules redistributing to the uropod. Reason: Polarization of the responding leukocyte is a downstream cellular event of CCR5 activation and part of the chemotactic response, not a distinct function of the chemokine. Retained as a non-core process consistent with chemokine biology. Supporting Evidence: PMID:9759849 Chemokines induce the formation of two specialized regions in the NK cell: the advancing front or leading edge, where chemokine receptors CCR2 and CCR5 cluster |
| GO:0007165 signal transduction | TAS PMID:10929056 Induction of interactions between CD44 and hyaluronic acid b... | MODIFY | Summary: Traceable author statement that MIP-1beta signals to T cells; the paper shows chemokine-triggered CD44 activation. Reason: Correct but uninformative. CCL4 initiates signaling specifically as an extracellular chemokine ligand of CCR5 (and of CCR1/CCR2b in its truncated form), so chemokine-mediated signaling pathway is the appropriate term; it is already supported by IBA. Proposed replacements: chemokine-mediated signaling pathway Supporting Evidence: PMID:8699119 MIP-1beta, and RANTES were all potent agonists for CC CKR5 |
| GO:0007165 signal transduction | TAS PMID:9558100 Dendritic cells express multiple chemokine receptors used as... | MODIFY | Summary: Traceable author statement associated with a study of chemokine receptors on dendritic cells used as HIV coreceptors. Reason: Signal transduction is too general for a chemokine ligand. The specific term chemokine-mediated signaling pathway describes what CCL4 does when it engages CCR5. Proposed replacements: chemokine-mediated signaling pathway Supporting Evidence: PMID:8699119 MIP-1beta, and RANTES were all potent agonists for CC CKR5 |
| GO:0007267 cell-cell signaling | TAS PMID:2521882 Mitogenic activation of human T cells induces two closely re... | ACCEPT | Summary: The pAT 744 (CCL4) cloning paper proposes it encodes a novel secreted lymphokine/cytokine produced by mitogen-activated T cells. Reason: As a secreted chemokine that acts on receptors of other cells, CCL4 mediates cell-cell signaling. General but correct. Supporting Evidence: PMID:2521882 suggest that pAT 464 and pAT 744 encode novel lymphokines/cytokines which may play roles during an immune response |
| GO:0008009 chemokine activity | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetic inference of chemokine activity across the CC chemokine family. CCL4 is a bona fide CC chemokine and potent CCR5 agonist. Reason: Chemokine activity is the core molecular function of CCL4; it is a secreted ligand that activates CCR5 with EC50 in the low nanomolar range and induces directed leukocyte migration. Supporting Evidence: PMID:8699119 MIP-1beta, and RANTES were all potent agonists for CC CKR5 file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. |
| GO:0008009 chemokine activity | IEA GO_REF:0000002 | ACCEPT | Summary: InterPro2GO mapping from chemokine domain signatures (IPR001811, IPR000827, IPR039809) to chemokine activity. Reason: Correct and at the right level of specificity for CCL4; duplicates the IBA and TAS annotations for the same term. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt SIMILARITY: Belongs to the intercrine beta (chemokine CC) family. |
| GO:0008009 chemokine activity | TAS PMID:10929056 Induction of interactions between CD44 and hyaluronic acid b... | ACCEPT | Summary: MIP-1beta used as a representative pro-inflammatory chemokine acting on human T cells. Reason: Correct core molecular function; duplicate of the IBA/IEA rows for the same term. Supporting Evidence: PMID:10929056 chemokines (e.g. MIP-1beta, interleukin-8, and RANTES) |
| GO:0009615 response to virus | TAS PMID:9743377 CXCR4 and CCR5 on human thymocytes: biological function and ... | MODIFY | Summary: In human thymocytes, productive infection by the CCR5-tropic HIV-1 strain Ba-L was specifically inhibited by MIP-1beta acting as a CCR5 agonist. Reason: The cited evidence is not a generic cellular response to virus but CCL4 actively restricting viral entry by occupying and down-modulating the CCR5 coreceptor. Defense response to virus is the accurate and more informative term. Proposed replacements: defense response to virus Supporting Evidence: PMID:9743377 this infection was specifically inhibited with the CCR5 agonist, macrophage inflammatory protein-1beta PMID:8525373 The chemokines RANTES, MIP-1 alpha, and MIP-1 beta were identified as the major HIV-SF produced by CD8+ T cells. |
| GO:0009636 response to toxic substance | IDA PMID:10841574 In vitro correlates of HIV-2-mediated HIV-1 protection. | UNDECIDED | Summary: Annotation from a study of HIV-2-mediated protection against HIV-1, where high beta-chemokine levels correlated with low HIV-1 replication and antibodies to beta-chemokines abrogated the resistance. The same term was assigned from this paper to CCL3 and CCL5. Reason: The cached record for this paper is abstract-only, and the abstract says nothing about a toxic substance; the relationship between CCL4 and a virus is not naturally expressed by response to toxic substance. However this is an experimental (IDA) annotation whose full text the curator read and this reviewer could not obtain (PMC full text returned empty), so it is left undecided rather than removed. If the intended claim is antiviral protection, a term such as GO:0051607 defense response to virus or GO:0046597 host-mediated suppression of symbiont invasion would be more apt. Supporting Evidence: PMID:10841574 High levels of beta-chemokines, the natural ligands of the CCR5 coreceptor, were correlated with low levels of viral replication, and resistance was abrogated by antibodies to beta-chemokines. |
| GO:0030335 positive regulation of cell migration | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetically inferred positive regulation of cell migration across the chemokine family; CCL4 itself appears in the WITH/FROM list, reflecting its own experimental grounding as a leukocyte chemoattractant. Reason: Promoting directed migration of CCR5-expressing leukocytes (monocytes, T cells, NK cells) is a core function of CCL4, and the node placement covers it directly. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. PMID:8699119 These data suggest that CC CKR5 is a G(i)-coupled receptor that may mediate monocyte responses to MIP-1alpha, MIP-1beta, and RANTES. |
| GO:0031726 CCR1 chemokine receptor binding | IPI PMID:7545673 Monocyte chemotactic protein-3 (MCP3) interacts with multipl... | KEEP AS NON CORE | Summary: Binding study on CCR1 (C-C CKR1) transfectants. Full-length MIP-1beta competed only weakly for the shared CCR1 binding site in this assay, but the naturally secreted truncated form MIP-1beta(3-69) is a genuine CCR1 agonist. Reason: CCR1 binding is real for CCL4 but secondary. The full-length protein competes poorly at CCR1, while the physiological N-terminally truncated form MIP-1beta(3-69) acquires CCR1 (and CCR2b) agonism; UniProt records the same. Retained as a non-core receptor interaction alongside the core CCR5 binding. Supporting Evidence: PMID:12070155 unlike the full-length protein, it also triggers a Ca(2+) response via CCR1 and CCR2b file:human/CCL4/CCL4-uniprot.txt is also a ligand for CCR1 and CCR2 isoform B. |
| GO:0031730 CCR5 chemokine receptor binding | IPI PMID:10383387 Extracellular cysteines of CCR5 are required for chemokine b... | ACCEPT | Summary: Mutagenesis of the four extracellular cysteines of CCR5 abolished MIP-1beta binding, directly demonstrating a CCL4-CCR5 binding interaction and mapping its structural requirements on the receptor. Reason: CCR5 binding is the core molecular function of CCL4 and is directly supported by this binding assay. Supporting Evidence: PMID:10383387 All cysteine mutants were unable to bind detectable levels of MIP-1beta, and did not respond functionally to CCR5 agonists. |
| GO:0031730 CCR5 chemokine receptor binding | IPI PMID:10679098 Lipopolysaccharide inhibits HIV-1 infection of monocyte- der... | ACCEPT | Summary: Study of CCR5 turnover in human macrophages, contrasting LPS-driven down-regulation with the rapid receptor turnover caused by the natural ligands MIP-1alpha and MIP-1beta. Reason: CCL4 is one of the natural ligands of CCR5 and this paper uses it as such; duplicates the core CCR5-binding annotation. Supporting Evidence: PMID:10679098 differed from the rapid turnover of these chemokine receptors observed in response to two natural ligands, macrophage-inflammatory protein-1alpha and -1beta |
| GO:0042802 identical protein binding | IPI PMID:20959807 Polymerization of MIP-1 chemokine (CCL3 and CCL4) and cleara... | MODIFY | Summary: Crystal structures show that CCL4 (MIP-1beta) self-associates into rod-shaped double-helical polymers, a reversible polymerization that buries receptor-binding surfaces and protects the chemokine from degradation of its monomeric form by insulin-degrading enzyme. Reason: The self-interaction is a specific homo-oligomerization with a defined structural basis and a regulatory role in gradient formation, so the generic identical protein binding term understates it. Protein homodimerization activity is the available GO term matching the UniProt SUBUNIT statement that CCL4 forms homodimers (as well as heterodimers with CCL3). Proposed replacements: protein homodimerization activity Supporting Evidence: PMID:20959807 Our crystal structures reveal that MIP-1 aggregation is a polymerization process and human MIP-1Ξ± and MIP-1Ξ² form rod-shaped, double-helical polymers. file:human/CCL4/CCL4-uniprot.txt SUBUNIT: Homodimer and heterodimer of MIP-1-alpha(4-69) |
| GO:0043922 host-mediated suppression of viral transcription | IDA PMID:10841574 In vitro correlates of HIV-2-mediated HIV-1 protection. | UNDECIDED | Summary: Annotation from the HIV-2/HIV-1 cross-protection study; peripheral blood mononuclear cells from HIV-2-infected subjects resisted CCR5-dependent HIV-1 challenge, resistance correlated with beta-chemokine levels and was abrogated by anti-beta-chemokine antibodies. The same term was assigned from this paper to CCL3 and CCL5. Reason: The established mechanism by which CCL4 restricts HIV-1 is at the level of entry, by occupying and down-modulating the CCR5 coreceptor, rather than by interfering with viral transcription, so the term appears to sit at the wrong step of the viral life cycle. Because the record cached here is abstract-only and the curator made an experimental (IDA) call from the full text, this is left undecided rather than modified or removed; GO:0046597 host-mediated suppression of symbiont invasion would be the mechanistically accurate alternative if the evidence is entry blockade. Supporting Evidence: PMID:10841574 we found that a significant proportion of peripheral blood mononuclear cells obtained from HIV-2-infected subjects resisted in vitro challenge with CCR5-dependent HIV-1 viruses but not CXCR4-dependent viruses PMID:12070155 MIP-1 beta(3-69) retains the abilities to induce down-modulation of surface expression of the chemokine receptor CCR5 and to inhibit the CCR5-mediated entry of HIV-1 in T cells. |
| GO:0048020 CCR chemokine receptor binding | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetically inferred CCR chemokine receptor binding across the CC chemokine family; CCL4 appears in the WITH/FROM list, reflecting its own experimental grounding as a CCR5 ligand. Reason: CCL4 binds CCR5 (core) and, in its truncated form, CCR1 and CCR2b. CCR chemokine receptor binding is the correct parent term for these activities. Supporting Evidence: PMID:10383387 All cysteine mutants were unable to bind detectable levels of MIP-1beta, and did not respond functionally to CCR5 agonists. file:human/CCL4/CCL4-uniprot.txt is also a ligand for CCR1 and CCR2 isoform B. |
| GO:0048245 eosinophil chemotaxis | IBA GO_REF:0000033 | MARK AS OVER ANNOTATED | Summary: Eosinophil chemotaxis propagated from an ancestral node (PTN008532501) whose extant experimental support comes from the eotaxin/CCR3 branch of the family (CCL24, CCL11, CCL7, CCL13, CCL5). CCL4 is not a CCR3 agonist, and CCR5 - its principal receptor - is not expressed on eosinophils. Reason: The node assertion fits the CCR3-binding eotaxins that seed it, but CCL4 has diverged in receptor usage within the family; its receptors are CCR5 and, for the truncated form, CCR1/CCR2b. CCR5 mRNA is absent from primary eosinophils, so a direct eosinophil-chemotactic role for CCL4 is not supported. Retained rather than removed because partial, indirect eosinophil recruitment through CCR1 cannot be formally excluded, but it is not a function of CCL4 in the sense the term asserts. Propagation Review Root cause: PROPAGATION BAD Failure modes: FUNCTIONAL DIVERGENCE CONTEXT OR TISSUE MISMATCH Sources checked: PANTHER:PTN008532501 SUPPORTS SOURCE BUT NOT TARGET The ancestral node carries eosinophil chemotaxis, but the extant experimental evidence under it comes from CCR3-binding eotaxins; CCL4's receptor repertoire (CCR5, and CCR1/CCR2b for the truncated form) does not include CCR3, and eosinophils do not express CCR5. Supporting Evidence: PMID:8699119 CC CKR5 mRNA was detected constitutively in primary adherent monocytes but not in primary neutrophils or eosinophils. file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. Binds |
| GO:0050850 positive regulation of calcium-mediated signaling | IMP PMID:10679098 Lipopolysaccharide inhibits HIV-1 infection of monocyte- der... | ACCEPT | Summary: In human macrophages, the natural CCR5 ligands MIP-1alpha and MIP-1beta drive rapid chemokine-receptor turnover, and CCR5 engagement by these ligands triggers Gi-coupled calcium-mediated signaling. Reason: Activation of calcium-mediated signaling in target cells is the immediate consequence of CCL4 binding CCR5 and is directly demonstrated for CCL4 in CCR5 transfectants, where the response is pertussis-toxin sensitive. Consistent with the core chemokine-signaling function. Supporting Evidence: PMID:10679098 differed from the rapid turnover of these chemokine receptors observed in response to two natural ligands, macrophage-inflammatory protein-1alpha and -1beta PMID:8699119 The calcium flux responses were completely blocked by treatment of transfected cells with pertussis toxin. |
| GO:0051928 positive regulation of calcium ion transport | IDA PMID:8699119 Cloning and functional expression of CC CKR5, a human monocy... | ACCEPT | Summary: MIP-1beta was a potent agonist for CCR5 in transfected HEK 293 cells as measured by calcium flux, with responses abolished by pertussis toxin, indicating Gi-coupled mobilization of calcium. Reason: Directly supported experimental annotation; increasing cytosolic calcium in CCR5-expressing cells is part of the core signaling output of CCL4. Supporting Evidence: PMID:8699119 MIP-1beta, and RANTES were all potent agonists for CC CKR5 (EC50 = 3-30 nM) when calcium flux was measured in transfected HEK 293 cells |
| GO:0061844 antimicrobial humoral immune response mediated by antimicrobial peptide | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: Antimicrobial peptide-mediated humoral immune response propagated across the CC chemokine family. Several CC chemokines have direct microbicidal activity in vitro, but no such activity has been reported specifically for CCL4. Reason: The node assertion is plausible for a small cationic chemokine, and the review does not have target-specific evidence against it, but CCL4 is characterized as a CCR5 agonist and leukocyte chemoattractant rather than as a direct antimicrobial effector. Retained as a non-core, phylogenetically-inferred function. Supporting Evidence: file:human/CCL4/CCL4-uniprot.txt Monokine with inflammatory and chemokinetic properties. Binds |
| GO:0070098 chemokine-mediated signaling pathway | IBA GO_REF:0000033 | ACCEPT | Summary: Phylogenetically inferred participation in the chemokine-mediated signaling pathway, as the ligand that initiates it. Reason: CCL4 initiates the chemokine signaling cascade by binding CCR5, which couples to Gi and mobilizes calcium. This is a core process for CCL4. Supporting Evidence: PMID:8699119 These data suggest that CC CKR5 is a G(i)-coupled receptor that may mediate monocyte responses to MIP-1alpha, MIP-1beta, and RANTES. |
| GO:2000503 positive regulation of natural killer cell chemotaxis | IDA PMID:7545673 Monocyte chemotactic protein-3 (MCP3) interacts with multipl... | ACCEPT | Summary: Chemokine binding and migration assays in the NK-like YT cell line and in CCR1 transfectants derived from it. Independent work shows that chemokines acting through CCR5, including MIP-1beta, recruit and polarize NK cells. Reason: NK cells express CCR5 and are recruited by MIP-1beta, so positive regulation of natural killer cell chemotaxis is a function CCL4 performs directly as the chemoattractant ligand. The same term is annotated to CCL3 and CCL5. Accepted, with the caveat that in the cited CCR1-transfectant assay full-length MIP-1beta was the weaker competitor; the curator read the full text and the function is independently supported. Supporting Evidence: PMID:9759849 Chemokines induce the formation of two specialized regions in the NK cell: the advancing front or leading edge, where chemokine receptors CCR2 and CCR5 cluster, which might guide the cells toward the chemotactic source PMID:7545673 All three C-C chemokines were able to cross-compete for binding sites on YT4/293 cells and induced directional migration of YT4/293 cells in vitro |
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Download this section (compressed HTML)Q: Does the IDA evidence in PMID:10841574 that produced the GO:0043922 (host-mediated suppression of viral transcription) and GO:0009636 (response to toxic substance) annotations for CCL3, CCL4 and CCL5 actually concern a transcriptional step, or is it coreceptor-level entry blockade that would be better captured by GO:0051607 or GO:0046597? The cached record is abstract-only, so these rows were left UNDECIDED.
Q: Should the receptor-binding and signaling annotations of CCL4 be split by proteoform? Full-length CCL4 is essentially CCR5-selective, whereas the predominant secreted form MIP-1-beta(3-69) also signals through CCR1 and CCR2b, so some receptor annotations arguably belong to the processed chain PRO_0000005165 rather than to the gene product as a whole.
Q: Is there a GO term, or should one exist, for ligand-induced down-modulation of a chemokine receptor as a distinct regulatory output of chemokine action? This is the mechanistic basis of CCL4's antiviral activity but is currently only captured indirectly.
Q: Does CCL4 have any direct microbicidal activity of its own, as the family-level IBA for GO:0061844 implies, or is that node assertion carried by other CC chemokines?
Experiment: Compare CCL4 treatment of primary CD4+ T cells challenged with R5-tropic HIV-1 against entry-independent readouts: use VSV-G-pseudotyped or CCR5-independent virus, or post-entry addition of CCL4, and quantify integrated provirus and LTR-driven transcription (e.g. nascent viral RNA) separately from entry measured by fusion reporter assays. Include a CCR5 internalization-deficient receptor mutant to test whether down-modulation rather than simple competition is required.
Hypothesis: The antiviral activity of CCL4 against R5-tropic HIV-1 is entirely accounted for by CCR5 occupancy and internalization, with no effect on steps after entry such as viral transcription.
Type: functional assay
Experiment: Assay purified human eosinophils in transwell and real-time chemotaxis assays against a CCL4 dose range, with CCL11 as a positive CCR3 control and CCL3 as a CCR1 comparator, and confirm receptor expression by flow cytometry for CCR1, CCR3 and CCR5. A negative result would justify removing the propagated eosinophil chemotaxis annotation.
Hypothesis: Eosinophils do not respond chemotactically to CCL4, contrary to the family-level GO:0048245 inference.
Type: chemotaxis assay
Experiment: Generate structure-guided depolymerizing mutants of CCL4 using the double-helical polymer interfaces defined crystallographically, verify oligomeric state by SEC and analytical ultracentrifugation, then compare monomer and polymer-competent forms for CCR5 binding, calcium flux, monocyte arrest on activated endothelium, susceptibility to insulin-degrading enzyme, and leukocyte recruitment in vivo.
Hypothesis: Polymerization of CCL4 regulates its in vivo chemotactic potency and half-life, as has been shown for CCL3.
Type: structure-function analysis
Experiment: Produce both proteoforms homogeneously, confirm the N-terminus by mass spectrometry, and compare calcium flux, beta-arrestin recruitment and chemotaxis in primary monocytes and T cells with selective CCR1, CCR2 and CCR5 antagonists, to establish which annotations belong to which proteoform.
Hypothesis: Receptor usage differs between full-length CCL4 and the natural MIP-1-beta(3-69) form in primary cells, not only in transfectants.
Type: receptor pharmacology
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