dagA

Existing Annotations Review

GO Term	Evidence	Action	Reason
GO:0005886 plasma membrane	IBA GO_REF:0000033	ACCEPT	Summary: CRAC is recruited from the cytosol to the plasma membrane upon chemoattractant stimulation, where its PH domain engages PIP3. This is a well-supported and functionally central localization. Reason: Regulated translocation of CRAC to the plasma membrane (the inner leaflet, where PIP3 accumulates) is directly documented and is where CRAC exerts its adenylyl cyclase-activating function. The phylogenetic (IBA) inference agrees with direct experimental evidence. Supporting Evidence: PMID:16267269 CRAC is rapidly and transiently recruited to the plasma membrane around the entire periphery of the cell
GO:0005543 phospholipid binding	IBA GO_REF:0000033	MODIFY	Summary: CRAC binds phosphoinositides (PI(3,4,5)P3 and PI(3,4)P2) via its PH domain. The general term phospholipid binding is correct but less informative than the specific 3-phosphoinositide binding terms, which are independently annotated with direct evidence. Reason: The specific lipid ligands of CRAC are the PI3K products PIP3 and PI(3,4)P2. Replacing the generic parent with the specific PIP3-binding term better captures the molecular function established experimentally. Propagation Review Root cause: TERM SCOPING PROBLEM Failure modes: GRANULARITY MISMATCH Sources checked: PANTHER:PTN008555303 · PANTHER node for PH-domain 3-phosphoinositide-binding adaptor proteins SUPPORTS SOURCE BUT NOT TARGET Generic phospholipid binding is correct but less informative than the specific PIP3 and PI(3,4)P2 binding established experimentally for CRAC UniProtKB:Q9UN19 · human DAPP1, a PIP3-binding PH-domain adaptor SUPPORTS SOURCE BUT NOT TARGET Source proteins bind 3-phosphoinositides; the specific PIP3-binding child term is preferable to the generic parent Proposed replacements: phosphatidylinositol-3,4,5-trisphosphate binding Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0043325 phosphatidylinositol-3,4-bisphosphate binding	IBA GO_REF:0000033	ACCEPT	Summary: The CRAC PH domain binds PI(3,4)P2 as well as PI(3,4,5)P3. This specific lipid-binding activity is directly supported. Reason: PI(3,4)P2 is one of the two PI3K products bound by the CRAC PH domain; the IBA inference matches the experimental characterization of CRAC as a 3-phosphoinositide-binding protein. Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0005737 cytoplasm	IEA GO_REF:0000044	ACCEPT	Summary: In unstimulated cells CRAC is a cytosolic protein, translocating to membranes only transiently upon stimulation. Cytoplasmic localization is correct. Reason: CRAC was originally purified and defined as a cytosolic regulator; its resting distribution is cytoplasmic, consistent with the UniProt subcellular location mapping. Supporting Evidence: PMID:8089184 A cytosolic protein that activates adenylyl cyclase, CRAC
GO:0046688 response to copper ion	HDA PMID:40863941 Proteomic Analysis of Heavy Metal-Induced Toxicity Using the...	MARK AS OVER ANNOTATED	Summary: This annotation derives from a 2D-gel proteomic screen in which the CRAC protein spot disappeared after copper exposure. This is a correlative change in protein abundance, not evidence that CRAC functions in a copper response. Reason: The proteomic study shows that CRAC abundance is reduced under copper stress (alongside formin-1), which is interpreted as copper disrupting aggregation-related proteins. It does not demonstrate that CRAC acts upstream of or within a physiological response to copper; the causal direction is the reverse (copper affects CRAC). Retained but flagged as over-annotation. Supporting Evidence: PMID:40863941 spot a corresponded to formin-1, spot b corresponded to CRAC PMID:40863941 Three spots (designated spots a, b, and c) were missing after Cu exposure
GO:0005829 cytosol	IDA PMID:15470246 A cell number counting factor regulates Akt/protein kinase B...	ACCEPT	Summary: Cell-fractionation and GFP-CRAC imaging assays in this study monitor the cytosol-to-membrane translocation of CRAC, consistent with a cytosolic resting pool. Reason: The study directly assays CRAC distribution between cytosol and membrane fractions, confirming the cytosolic localization of resting CRAC. Supporting Evidence: PMID:15470246 there is a quantitative change in the translocation of CRAC in the presence of anti-countin antibodies or recombinant countin
GO:0005829 cytosol	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: Live-cell imaging shows PH(Crac)-GFP returning to the cytosol after membrane translocation (adaptation), documenting the cytosolic pool. Reason: Quantitative single-cell imaging of PH(Crac)-GFP directly demonstrates cytosolic localization in the basal/adapted state. Supporting Evidence: PMID:15563608 PH domain-containing proteins evenly translocate to the plasma membrane and then quickly return to the cytosol
GO:0005829 cytosol	IDA PMID:16267269 Phosphoinositide 3-kinase activity controls the chemoattract...	ACCEPT	Summary: CRAC is defined and used here as a cytosolic (PH-domain) regulator that is recruited to membranes only on stimulation. Reason: The paper directly studies CRAC recruitment dynamics and identifies it as the cytosolic regulator of adenylyl cyclase, supporting cytosolic localization. Supporting Evidence: PMID:16267269 the pleckstrin homology (PH) domain-containing protein cytosolic regulator of adenylyl cyclase (CRAC)
GO:0005829 cytosol	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC is a cytosolic protein recruited to the membrane during chemoattractant signaling; this study established the imaging paradigm for that translocation. Reason: Directly supports the cytosolic nature of CRAC, whose recruitment from the cytosol reflects G protein activation. Supporting Evidence: PMID:9778249 transmitted by the recruitment of cytosolic proteins
GO:0005886 plasma membrane	IDA PMID:16267269 Phosphoinositide 3-kinase activity controls the chemoattract...	ACCEPT	Summary: Direct imaging shows CRAC recruited to the plasma membrane periphery upon chemoattractant stimulation. Reason: This is a core, directly observed localization where CRAC engages PIP3 and functions in ACA activation. Supporting Evidence: PMID:16267269 CRAC is rapidly and transiently recruited to the plasma membrane around the entire periphery of the cell
GO:0005886 plasma membrane	IDA PMID:23132928 Delineating the core regulatory elements crucial for directe...	ACCEPT	Summary: The PH(Crac) biosensor reports plasma-membrane phosphoinositides, confirming CRAC recruitment to the plasma membrane. Reason: PH(Crac)-GFP measures plasma-membrane PIP3/PI(3,4)P2, directly placing CRAC at the plasma membrane during chemotactic responses. Supporting Evidence: PMID:23132928 a biosensor that measures increased levels of these plasma membrane (PM) phosphoinositides
GO:0005886 plasma membrane	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocates to binding sites on the inner face of the plasma membrane upon G protein activation. Reason: Directly documented recruitment of CRAC to the plasma membrane (inner leaflet), a core functional localization. Supporting Evidence: PMID:9778249 The translocation of the PH domain-containing protein CRAC in D. discoideum to binding sites on the inner face of the plasma membrane
GO:0006935 chemotaxis	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: Mutational analysis shows CRAC is required for chemotaxis in addition to its role in ACA activation; a CRAC mutant unable to bind PI3K products fails to support chemotaxis. Reason: Directly demonstrated by CRAC mutants; PI3K-dependent CRAC function is required for directed migration, a core biological role. Supporting Evidence: PMID:15668169 in addition to its essential role in the activation of ACA, CRAC is involved in regulating chemotaxis PMID:15668169 A CRAC mutant that has lost the capacity to bind PI3K products does not support chemotaxis and shows minimal ACA activation
GO:0006935 chemotaxis	IMP PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocation to the leading edge is a hallmark of the chemotactic response; this study established leading-edge recruitment of CRAC in chemotaxing cells. Reason: The gene product's leading-edge localization during chemotaxis supports a role in directed migration. Supporting Evidence: PMID:9778249 G protein signaling events are activated at the leading edge of chemotactic cells
GO:0008047 enzyme activator activity	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	ACCEPT	Summary: CRAC is required for receptor- and G protein-mediated activation of adenylyl cyclase, acting as an adapter linking free Gbetagamma to ACA activation. This is the defining core molecular function of CRAC. Reason: The founding genetic/biochemical study shows dagA/CRAC is essential for adenylyl cyclase activation and proposes it connects Gbetagamma to the cyclase, i.e. an enzyme (adenylyl cyclase) activator role. Supporting Evidence: PMID:8089184 is required for receptor and G protein-mediated activation of adenylyl cyclase in PMID:8089184 CRAC acts to connect free G protein beta gamma subunits to adenylyl cyclase activation
GO:0009898 cytoplasmic side of plasma membrane	IDA PMID:15161938 Sensitization of Dictyostelium chemotaxis by phosphoinositid...	ACCEPT	Summary: The CRAC PH domain binds the cytoplasmic (inner) face of the plasma membrane, forming PH(Crac)-GFP patches that mark PIP3 at the leading edge. Reason: PH(Crac)-GFP localizes to inner-leaflet membrane patches, consistent with the cytoplasmic side of the plasma membrane. Supporting Evidence: PMID:15161938 translocation of PH(Crac)-GFP from the cytosol to multiple patches
GO:0016020 membrane	IDA PMID:15470246 A cell number counting factor regulates Akt/protein kinase B...	MODIFY	Summary: CRAC translocates to the membrane fraction; the general term membrane is less informative than plasma membrane, which is the actual site of CRAC recruitment. Reason: The membrane to which CRAC translocates is the plasma membrane (inner leaflet). Replacing the generic term with plasma membrane improves specificity while remaining supported by the fractionation data. Proposed replacements: plasma membrane Supporting Evidence: PMID:15470246 CRAC translocation to membranes is required for the GTPγS stimulation of adenylyl cyclase activity
GO:0016020 membrane	IDA PMID:7790361 Chemoattractant and GTP gamma S-mediated stimulation of aden...	MODIFY	Summary: Chemoattractant stimulation drives translocation of CRAC from the cytosolic to the membrane fraction. The specific compartment is the plasma membrane. Reason: The biochemical membrane fraction that CRAC translocates to corresponds to the plasma membrane, where guanine-nucleotide-generated Gbetagamma binding sites reside. Plasma membrane is the more informative term. Proposed replacements: plasma membrane Supporting Evidence: PMID:7790361 chemoattractants promote translocation of CRAC from the cytosolic to the membrane fraction
GO:0031143 pseudopodium	IDA PMID:23132928 Delineating the core regulatory elements crucial for directe...	ACCEPT	Summary: CRAC (via PH(Crac)) is among the proteins enriched in pseudopods during chemotaxis, reflecting leading-edge PIP3 accumulation. Reason: Pseudopod enrichment of PIP3-binding proteins such as CRAC is directly observed during folic-acid- and cAMP-mediated chemotaxis. Supporting Evidence: PMID:23132928 Proteins enriched in the pseudopods during chemotaxis also relocalize transiently to the plasma membrane during uniform FA stimulation
GO:0031252 cell leading edge	IDA PMID:17126332 Time-resolved responses to chemoattractant, characteristic o...	ACCEPT	Summary: CRAC accumulates at the leading edge of chemotaxing cells, marking sites of PIP3 production during front responses to chemoattractant. Reason: Leading-edge localization of CRAC/PH(Crac) is a well-established, directly imaged feature of the chemotactic response. Supporting Evidence: PMID:23132928 at the leading edge of migrating cells can be identified by the plekstrin homology (PH) domain of the cytosolic regulator of adenylyl cyclase (CRAC)
GO:0031256 leading edge membrane	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: PH(Crac)-GFP forms a highly polarized crescent at the leading-front plasma membrane in a chemoattractant gradient. Reason: Single-cell imaging directly shows CRAC translocating to the plasma membrane at the leading front, i.e. the leading edge membrane. Supporting Evidence: PMID:15563608 PH domain-containing proteins translocate from the cytosol to the plasma membrane at the leading front
GO:0031256 leading edge membrane	IDA PMID:9778249 G protein signaling events are activated at the leading edge...	ACCEPT	Summary: CRAC translocates to inner-face plasma membrane binding sites selectively at the stimulated (leading) edge of chemotactic cells. Reason: Directly documented leading-edge plasma-membrane recruitment of CRAC. Supporting Evidence: PMID:9778249 The translocation of the PH domain-containing protein CRAC in D. discoideum to binding sites on the inner face of the plasma membrane
GO:0106070 regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: CRAC is essential for chemoattractant-mediated activation of the aggregation adenylyl cyclase (ACA), placing it within the regulation of the adenylate cyclase-activating GPCR signaling pathway. Reason: A defining, directly demonstrated role in which CRAC is required for GPCR/PI3K-dependent activation of ACA. Supporting Evidence: PMID:15668169 CRAC is essential for the chemoattractant-mediated activation of the adenylyl cyclase ACA
GO:0106070 regulation of adenylate cyclase-activating G protein-coupled receptor signaling pathway	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	ACCEPT	Summary: The founding study shows dagA/CRAC is required for receptor and G protein-mediated activation of adenylyl cyclase. Reason: Genetic disruption of dagA abolishes receptor/G protein activation of adenylyl cyclase, a core function within this pathway. Supporting Evidence: PMID:8089184 is required for receptor and G protein-mediated activation of adenylyl cyclase in
GO:0000165 MAPK cascade	IMP PMID:9020088 The Dictyostelium mitogen-activated protein kinase ERK2 is r...	KEEP AS NON CORE	Summary: CRAC is required for proper adaptation of the MAP kinase ERK2, linking it to MAPK signaling. This is an indirect effect via cAMP/ACA signaling rather than a core CRAC function. Reason: Loss of CRAC alters ERK2 adaptation kinetics, so CRAC is functionally upstream of the ERK2 (MAPK) response. However, this is a downstream consequence of CRAC's role in cAMP signaling rather than a distinct core molecular function; retained as non-core. Supporting Evidence: PMID:9020088 CRAC, a PH domain-containing protein required for adenylyl cyclase activation, is also required for proper ERK2 adaptation
GO:0019887 protein kinase regulator activity	IMP PMID:9020088 The Dictyostelium mitogen-activated protein kinase ERK2 is r...	MARK AS OVER ANNOTATED	Summary: This molecular-function annotation is inferred from CRAC being required for ERK2 adaptation. CRAC has no known direct kinase-regulatory activity; the effect on ERK2 is indirect through cAMP signaling. Reason: CRAC is a PIP3-binding adapter, not a direct regulator of a protein kinase. The requirement for CRAC in ERK2 adaptation is mediated through its role in the cAMP/adenylyl cyclase pathway, so assigning a direct protein kinase regulator activity over-interprets the phenotype. Supporting Evidence: PMID:9020088 CRAC, a PH domain-containing protein required for adenylyl cyclase activation, is also required for proper ERK2 adaptation
GO:0001891 phagocytic cup	IDA PMID:16968738 Selective membrane exclusion in phagocytic and macropinocyti...	UNDECIDED	Summary: The cited study characterizes selective membrane sorting at phagocytic and macropinocytic cups. The cached record is abstract-only and does not mention CRAC, so the specific localization of CRAC cannot be verified here. Reason: Per policy, this experimental (IDA) curator annotation is not overruled. The cached abstract does not report CRAC, and no full text is available to confirm the CRAC-specific phagocytic-cup localization, so the annotation is left undecided pending access to the primary evidence.
GO:0070685 macropinocytic cup	IDA PMID:16968738 Selective membrane exclusion in phagocytic and macropinocyti...	UNDECIDED	Summary: As above, this study addresses membrane sorting at macropinocytic cups; the cached abstract-only record does not mention CRAC, so CRAC-specific localization cannot be verified. Reason: Cannot verify the CRAC-specific macropinocytic-cup localization from the abstract-only cached publication; the experimental annotation is retained as undecided rather than overruled.
GO:0030587 sorocarp development	HMP PMID:17659086 High-throughput analysis of spatio-temporal dynamics in Dict...	KEEP AS NON CORE	Summary: dagA-null cells are developmentally defective (developmentally null cluster). Sorocarp (fruiting body) development is disrupted because CRAC-dependent ACA activation is required for aggregation, but this is a broad downstream developmental outcome rather than CRAC's core function. Reason: High-throughput phenotyping places dagA among developmentally null mutants, so it acts within sorocarp development. This reflects the developmental consequence of losing cAMP signaling rather than a distinct molecular function; retained as non-core. Supporting Evidence: PMID:17659086 mutants in which genes such as mkpA, piaA, yakA and dagA are disrupted
GO:0007188 adenylate cyclase-modulating G protein-coupled receptor signaling pathway	IDA PMID:17606871 Locally controlled inhibitory mechanisms are involved in euk...	ACCEPT	Summary: CRAC (PH(Crac)-GFP) reports PIP3 dynamics downstream of GPCR activation and is a component of the adenylate cyclase-modulating GPCR signaling pathway. Reason: Directly observed CRAC/PIP3 responses during GPCR-mediated chemosensing support involvement in this signaling pathway. Supporting Evidence: PMID:17606871 The gradients induce a stable accumulation of the PIP3 reporter PHCrac-GFP in the front of cells near the cAMP source
GO:0043325 phosphatidylinositol-3,4-bisphosphate binding	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: CRAC binds the PI3K products PI(3,4)P2 and PI(3,4,5)P3; mutants unable to bind these lipids lose function. Reason: Directly supported lipid-binding molecular function of the CRAC PH domain; PI(3,4)P2 is one of the ligands. Supporting Evidence: PMID:15668169 pleckstrin homology (PH) domain-containing proteins that bind to the PI3K products PI(3,4)P2 and PI(3,4,5)P3, such as CRAC
GO:0048015 phosphatidylinositol-mediated signaling	TAS PMID:15366706 Chemotaxis: signalling the way forward.	KEEP AS NON CORE	Summary: CRAC is a downstream effector of PI3K-generated 3-phosphoinositides, binding them via its PH domain to transduce chemoattractant signals. It thus acts within phosphatidylinositol-mediated signaling. Reason: CRAC operates within the PI3K/PIP3 signaling branch, but the more specific and central descriptions of its role are its PIP3 binding and adenylyl cyclase activation. Retained as non-core context. Supporting Evidence: PMID:16267269 leading to the production of 3-phosphoinositides (3-PI) to which CRAC binds via its PH domain
GO:0030435 sporulation resulting in formation of a cellular spore	TAS PMID:15473840 Chemoattractant signaling in dictyostelium discoideum.	KEEP AS NON CORE	Summary: Because dagA-null cells fail to complete development, spore formation is indirectly affected. This is a distal developmental consequence of loss of CRAC-dependent aggregation signaling. Reason: dagA is among developmentally null mutants; sporulation failure follows from the block in early cAMP signaling/aggregation rather than from a direct role of CRAC in sporulation. Retained as non-core. Supporting Evidence: PMID:17659086 mutants in which genes such as mkpA, piaA, yakA and dagA are disrupted
GO:0005547 phosphatidylinositol-3,4,5-trisphosphate binding	IDA PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	ACCEPT	Summary: The CRAC PH domain binds PI(3,4,5)P3 (PIP3); a CRAC mutant that cannot bind PI3K products loses both ACA activation and chemotaxis. This is a core molecular function. Reason: Direct evidence that PIP3 binding by CRAC is required for its function; this is the primary molecular activity of the protein. Supporting Evidence: PMID:15668169 A CRAC mutant that has lost the capacity to bind PI3K products does not support chemotaxis and shows minimal ACA activation
GO:0008064 regulation of actin polymerization or depolymerization	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	KEEP AS NON CORE	Summary: CRAC, like Akt/PKB, contributes to regulation of actin polymerization during chemotaxis downstream of PIP3. This is a secondary role relative to ACA activation. Reason: PIP3-binding effectors including CRAC influence actin dynamics during directed migration, but this is one downstream branch of CRAC signaling rather than its defining function; retained as non-core. Supporting Evidence: PMID:17606871 Both CRAC and Akt/PKB play roles in the regulation of actin polymerization during chemotaxis
GO:0031152 aggregation involved in sorocarp development	IMP PMID:15668169 The PI3K-mediated activation of CRAC independently regulates...	KEEP AS NON CORE	Summary: CRAC is essential for activation of ACA, which produces the cAMP that drives chemotactic aggregation. Loss of CRAC blocks normal aggregation. Reason: Aggregation is the developmental process that depends on CRAC-mediated cAMP relay; it is a downstream outcome of CRAC's core signaling function. Retained as non-core. Supporting Evidence: PMID:15668169 CRAC is essential for the chemoattractant-mediated activation of the adenylyl cyclase ACA, which converts ATP into cAMP, the primary chemoattractant for D. discoideum
GO:0031152 aggregation involved in sorocarp development	IMP PMID:8089184 CRAC, a cytosolic protein containing a pleckstrin homology d...	KEEP AS NON CORE	Summary: dagA is required for normal development; re-expression of the dagA cDNA restores normal development in mutant cells, consistent with a role in aggregation. Reason: Genetic evidence links dagA to aggregation-stage development via ACA activation; downstream developmental process, retained as non-core. Supporting Evidence: PMID:8089184 the cDNA restores normal development when constitutively expressed in
GO:0007188 adenylate cyclase-modulating G protein-coupled receptor signaling pathway	IDA PMID:15563608 Quantitative imaging of single live cells reveals spatiotemp...	ACCEPT	Summary: PH(Crac)-GFP translocation dynamics directly report GPCR/G protein activation and PIP3 production, situating CRAC in the adenylate cyclase-modulating GPCR signaling pathway. Reason: Quantitative imaging of CRAC translocation as a readout of GPCR signaling supports its involvement in this pathway. Supporting Evidence: PMID:15563608 monitoring the dynamics of PH(Crac)-GFP translocation in single living cells

Gene Description

Existing Annotations Review

Core Functions

CRAC is a cytosolic PH-domain adapter that, upon chemoattractant stimulation, binds the PI3K products PI(3,4,5)P3 and PI(3,4)P2 and translocates to the plasma membrane (leading-edge inner leaflet), coupling GPCR/PI3K signaling to downstream effectors.

CRAC is required for receptor- and G protein (Gbetagamma)-mediated activation of the aggregation-stage adenylyl cyclase (ACA), functioning as an activator that links activated G protein signaling to cAMP synthesis for intercellular signal relay.

References

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