Tes (testin) is a cytoplasmic LIM-domain scaffold protein of the prickle/espinas/testin family, with an N-terminal PET domain and three C-terminal zinc-binding LIM domains. Its LIM domains bind cytoskeletal and focal adhesion proteins: LIM1 binds zyxin, which recruits Tes to focal adhesions; LIM3 binds the EVH1 domain of Mena (but not VASP or EVL), competing with FPPPP ligands and modulating Mena-dependent cell migration; and LIM2-3 binds the N-terminal extension of the testis actin-related protein Arp7A (ACTL7A). The N- and C-terminal halves associate in a closed conformation that masks partner binding. Tes localises to focal adhesions, actin stress fibres and cell-cell contacts, contributes to stress-fibre organisation and cell spreading, and in rat testis forms an Arp7A-Tes-Mena complex in the subacrosomal layer (acroplaxome) of round spermatids. Forced expression suppresses tumour-cell growth, and TES is a candidate tumour suppressor frequently silenced by promoter methylation.
| GO Term | Evidence | Action | Reason |
|---|---|---|---|
| GO:0005737 cytoplasm | IBA GO_REF:0000033 | KEEP AS NON CORE | Summary: Cytoplasmic localisation inherited across the prickle/espinas/testin (PET-LIM) family. Reason: PAINT placed cytoplasm at the family node with fly, frog, mouse and human Tes donors; rat Tes is a canonical member of that clade and nothing suggests loss. Tes is a soluble LIM-domain scaffold with no transmembrane segment or signal peptide; it distributes between a diffuse cytoplasmic pool, actin stress fibres and focal adhesions, and in round spermatids the subacrosomal layer. Cytoplasm is the umbrella location that covers this whole distribution; the specific site of action (focal adhesion) is captured by its own rows, so this row is kept as an accurate but peripheral location. Supporting Evidence: file:rat/Tes/Tes-uniprot.txt Belongs to the prickle / espinas / testin family. PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact |
| GO:0005737 cytoplasm | IEA GO_REF:0000120 | KEEP AS NON CORE | Summary: UniProt subcellular-location keyword mapping (Cytoplasm, by similarity) to cytoplasm. Reason: Tes is a soluble LIM-domain scaffold with no transmembrane segment or signal peptide; it distributes between a diffuse cytoplasmic pool, actin stress fibres and focal adhesions, and in round spermatids the subacrosomal layer. Cytoplasm is the umbrella location that covers this whole distribution; the specific site of action (focal adhesion) is captured by its own rows, so this row is kept as an accurate but peripheral location. Supporting Evidence: PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact |
| GO:0005737 cytoplasm | ISO GO_REF:0000121 | KEEP AS NON CORE | Summary: Transferred from mouse Tes, whose cytoplasm IDA comes from the spermatid immunostaining in PMID:21278383. Reason: Tes is a soluble LIM-domain scaffold with no transmembrane segment or signal peptide; it distributes between a diffuse cytoplasmic pool, actin stress fibres and focal adhesions, and in round spermatids the subacrosomal layer. Cytoplasm is the umbrella location that covers this whole distribution; the specific site of action (focal adhesion) is captured by its own rows, so this row is kept as an accurate but peripheral location. Supporting Evidence: PMID:21278383 all three proteins are actually localized in the subacrosomal layer that links the acrosome to the nuclear envelope PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact |
| GO:0005737 cytoplasm | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: UniProt similarity transfer of cytoplasm from mouse Tes (P47226). Reason: Tes is a soluble LIM-domain scaffold with no transmembrane segment or signal peptide; it distributes between a diffuse cytoplasmic pool, actin stress fibres and focal adhesions, and in round spermatids the subacrosomal layer. Cytoplasm is the umbrella location that covers this whole distribution; the specific site of action (focal adhesion) is captured by its own rows, so this row is kept as an accurate but peripheral location. Supporting Evidence: PMID:21278383 all three proteins are actually localized in the subacrosomal layer that links the acrosome to the nuclear envelope PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact |
| GO:0005829 cytosol | ISO GO_REF:0000121 | KEEP AS NON CORE | Summary: Cytosol transferred from human TES (IDA, Human Protein Atlas immunostaining). Reason: Outside adhesions Tes is a soluble cytosolic protein whose closed (N-/C-terminal self-associated) conformation regulates its zyxin-dependent recruitment to focal adhesions; the cytosolic pool is real but is the reservoir, not the site of function. Supporting Evidence: PMID:12695497 we demonstrate that the two halves of Tes interact with each other in vitro and in vivo |
| GO:0005925 focal adhesion | IEA GO_REF:0000044 | ACCEPT | Summary: UniProt subcellular location (Cell junction, focal adhesion) mapped to focal adhesion. Reason: Focal adhesion is the best-established site of action of Tes: human TES localises to focal adhesions and is recruited there by direct binding of LIM1 to zyxin. The rat protein is highly conserved and UniProt transfers this location by similarity. Supporting Evidence: PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact PMID:12695497 Tes is a component of focal adhesions that, when expressed, negatively regulates proliferation of T47D breast carcinoma cells. PMID:21278383 Tes is recruited to focal adhesions via a direct interaction between its LIM1 domain and zyxin (27). |
| GO:0005925 focal adhesion | ISO GO_REF:0000121 | ACCEPT | Summary: Focal adhesion transferred from human TES (IDA/HDA). Reason: Focal adhesion is the best-established site of action of Tes: human TES localises to focal adhesions and is recruited there by direct binding of LIM1 to zyxin. The rat protein is highly conserved and UniProt transfers this location by similarity. Supporting Evidence: PMID:12571287 the TES protein is localised at focal adhesions, actin stress fibres and areas of cell-cell contact PMID:12695497 Tes is a component of focal adhesions that, when expressed, negatively regulates proliferation of T47D breast carcinoma cells. PMID:21278383 Tes is recruited to focal adhesions via a direct interaction between its LIM1 domain and zyxin (27). |
| GO:0008270 zinc ion binding | IEA GO_REF:0000002 | KEEP AS NON CORE | Summary: InterPro2GO electronic annotation (via the IPR010442 PET domain entry) to zinc ion binding. Reason: Each of the three LIM domains is a double zinc finger; the crystal structure of the Tes LIM2-3 tandem contains two zinc ions per LIM domain. Zinc binding is structural (it folds the protein-interaction modules) rather than the functional activity of Tes, so it is kept as true but non-core. Supporting Evidence: PMID:21278383 Using a simple metric based on the four common zinc ions in the tandem LIM structures file:rat/Tes/Tes-uniprot.txt LIM zinc-binding 1 |
| GO:0008270 zinc ion binding | ISO GO_REF:0000121 | KEEP AS NON CORE | Summary: Zinc ion binding transferred from human TES (IDA, LIM2-3 crystal structure). Reason: Each of the three LIM domains is a double zinc finger; the crystal structure of the Tes LIM2-3 tandem contains two zinc ions per LIM domain. Zinc binding is structural (it folds the protein-interaction modules) rather than the functional activity of Tes, so it is kept as true but non-core. Supporting Evidence: PMID:21278383 Using a simple metric based on the four common zinc ions in the tandem LIM structures file:rat/Tes/Tes-uniprot.txt LIM zinc-binding 1 |
| GO:0008270 zinc ion binding | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: Zinc ion binding transferred by UniProt similarity from human TES. Reason: Each of the three LIM domains is a double zinc finger; the crystal structure of the Tes LIM2-3 tandem contains two zinc ions per LIM domain. Zinc binding is structural (it folds the protein-interaction modules) rather than the functional activity of Tes, so it is kept as true but non-core. Supporting Evidence: PMID:21278383 Using a simple metric based on the four common zinc ions in the tandem LIM structures file:rat/Tes/Tes-uniprot.txt LIM zinc-binding 1 |
| GO:0008285 negative regulation of cell population proliferation | ISO GO_REF:0000121 | KEEP AS NON CORE | Summary: Negative regulation of cell proliferation transferred from human TES (IMP, PMID:11420696). Reason: Forced expression of human TES suppresses colony formation of HeLa and OVCAR5 cells and growth of T47D cells, and TES is a candidate tumour suppressor silenced by promoter methylation. These are overexpression phenotypes in tumour lines with no defined mechanism, and no rat data exist; the term is retained as a supported but peripheral consequence of its adhesion/cytoskeletal scaffold role. Supporting Evidence: PMID:11420696 forced expression of TES in HeLa or OVCAR5 cells, resulted in a profound reduction in growth potential PMID:12695497 Tes is a component of focal adhesions that, when expressed, negatively regulates proliferation of T47D breast carcinoma cells. |
| GO:0008285 negative regulation of cell population proliferation | ISS GO_REF:0000024 | KEEP AS NON CORE | Summary: Negative regulation of cell proliferation transferred by UniProt similarity from human TES. Reason: Forced expression of human TES suppresses colony formation of HeLa and OVCAR5 cells and growth of T47D cells, and TES is a candidate tumour suppressor silenced by promoter methylation. These are overexpression phenotypes in tumour lines with no defined mechanism, and no rat data exist; the term is retained as a supported but peripheral consequence of its adhesion/cytoskeletal scaffold role. Supporting Evidence: PMID:11420696 forced expression of TES in HeLa or OVCAR5 cells, resulted in a profound reduction in growth potential PMID:12695497 Tes is a component of focal adhesions that, when expressed, negatively regulates proliferation of T47D breast carcinoma cells. |
| GO:0032991 protein-containing complex | ISO GO_REF:0000121 | KEEP AS NON CORE | Summary: Protein-containing complex transferred from human TES (IDA, Arp7A-Tes-Mena complex, PMID:21278383). Reason: The donor annotation is the Arp7A-Tes-Mena complex, and the same paper immunoprecipitated this complex from rat testis, so the claim holds directly for rat Tes. GO has no specific complex term for it, so the generic term is as precise as available; it is kept as a true but non-core location (the binding activity is captured as a NEW MF row). Supporting Evidence: PMID:21278383 Arp7A, Tes, and Mena are complexed to each other in rat testes extracts PMID:21278383 the N-terminal extension to the actin-like fold of Arp7A interacts directly with the LIM2 domain of Tes |
| GO:0008092 cytoskeletal protein binding | IPI PMID:21278383 Molecular recognition of the Tes LIM2-3 domains by the actin... | NEW | Summary: Proposed: Tes LIM domains bind cytoskeletal/focal-adhesion proteins (Mena EVH1 via LIM3, zyxin via LIM1, Arp7A via LIM2-3); the Tes-Mena-Arp7A complex was immunoprecipitated from rat testis. Reason: Tes has no GO molecular-function row other than structural zinc binding. Its established activity is binding to cytoskeletal proteins through its LIM domains: LIM3 binds the Mena EVH1 domain (crystal structure), LIM1 binds zyxin directly, LIM2-3 binds the Arp7A N-terminus (crystal structure), and Tes co-purifies with actin. In rat, Tes is complexed with Mena and Arp7A in testis extracts. This is the activity itself, not a phenotype, and is more informative than protein binding; it is not an ancestor or descendant of any term on the gene. Supporting Evidence: PMID:21278383 Arp7A, Tes, and Mena are complexed to each other in rat testes extracts PMID:18158903 the LIM3 domain of Tes, which lacks the FPPPP motif, binds to the EVH1 domain of Mena, but not to those of VASP or Evl PMID:21278383 the N-terminal extension to the actin-like fold of Arp7A interacts directly with the LIM2 domain of Tes PMID:21278383 Tes is recruited to focal adhesions via a direct interaction between its LIM1 domain and zyxin (27). PMID:12695497 Coimmunoprecipitations demonstrate that in vivo Tes is complexed with actin, Mena, and vasodilator-stimulated phosphoprotein (VASP). |
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Download this section (compressed HTML)Q: What is the function of the Arp7A-Tes-Mena complex in the rat acroplaxome, and does GO need a cellular-component term for the acroplaxome (subacrosomal layer) to capture this location?
Q: What signal opens the closed Tes conformation to allow zyxin binding and focal-adhesion recruitment, and is it conserved in rat?
Q: Is the growth-suppressive effect of TES overexpression a direct consequence of its focal-adhesion scaffolding (e.g. via Mena or RhoA), or an indirect effect in tumour lines?
Experiment: Germ-cell-specific Tes knockout in rat, followed by electron microscopy of acrosome-acroplaxome-nuclear envelope attachment and localisation of Arp7A and Mena.
Hypothesis: Tes is required for acroplaxome integrity and acrosome anchoring in rat spermatids.
Type: genetic knockout with ultrastructural analysis
Experiment: Express LIM3 point mutants that abolish Mena EVH1 binding in Tes-null rat fibroblasts and measure Mena localisation, lamellipodial dynamics and migration.
Hypothesis: Tes LIM3 limits Mena activity at focal adhesions in rat cells.
Type: structure-guided mutagenesis with live imaging
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