id: A0A0D2VGR4 gene_symbol: coWts aliases: - CAOG_00619 - CAOG_000619 - Warts - LATS product_type: PROTEIN status: DRAFT taxon: id: NCBITaxon:595528 label: Capsaspora owczarzaki ATCC 30864 description: >- coWts (CAOG_00619) is the single Warts/LATS-family (NDR subfamily, AGC group) serine/threonine protein kinase of the filasterean amoeba Capsaspora owczarzaki, a close unicellular relative of animals. The 1205-residue protein has a long, largely disordered N-terminal region, a LATS-type MOB-binding region, a C-terminal kinase domain with the conserved ATP-binding lysine and catalytic aspartate, and an AGC C-terminal extension carrying a predicted hydrophobic-motif phosphorylation site. In Capsaspora it acts in the Hippo pathway upstream of the Yorkie/YAP/TAZ ortholog coYki: cells lacking the coWts kinase domain show increased nuclear accumulation of coYki, more strongly than cells lacking the upstream kinase coHpo, consistent with cytoplasmic retention of coYki by Warts-dependent phosphorylation as in animals, although coWts kinase activity and coYki phosphorylation have not been measured directly. Loss of coWts does not increase proliferation (it slows growth in shaking culture) but produces a subpopulation of elongated cells that undergo a myosin-sensitive contractile behaviour, rescued by a coWts transgene, and within multicellular aggregates causes denser cell packing, reduced aggregate circularity and shorter filopodia. These phenotypes are mimicked by a hyperactive coYki that cannot be phosphorylated at its predicted Warts sites and require coYki's TEAD-binding residue, indicating that coWts shapes cytoskeletal behaviour and aggregate architecture by restraining coYki-dependent transcription. existing_annotations: - term: id: GO:0000166 label: nucleotide binding evidence_type: IEA original_reference_id: GO_REF:0000104 qualifier: enables supporting_entities: - UniRule:UR001503341 review: summary: >- UniRule-based assignment from the protein kinase ATP-binding signature. coWts has a canonical protein kinase domain (residues 711-1045) with the PROSITE ATP-binding motif and the conserved lysine (K740). action: KEEP_AS_NON_CORE reason: >- Correct but uninformative: nucleotide binding is the parent of the ATP binding annotation already present, and ATP binding is itself only a component of the kinase activity. Kept as a true, non-core statement; no experimental nucleotide binding data exist. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS00107; PROTEIN_KINASE_ATP; 1. - term: id: GO:0004672 label: protein kinase activity evidence_type: IEA original_reference_id: GO_REF:0000002 qualifier: enables supporting_entities: - InterPro:IPR008271 review: summary: >- InterPro2GO from the Ser/Thr kinase active-site signature (IPR008271). The catalytic loop aspartate (D834, proton acceptor) is annotated as present. action: KEEP_AS_NON_CORE reason: >- Correct, but the more specific protein serine/threonine kinase activity term is also present and is the better description of a Warts/LATS AGC kinase. Kept as a true, less specific parent. Activity has not been assayed for coWts. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS00108; PROTEIN_KINASE_ST; 1. - term: id: GO:0004674 label: protein serine/threonine kinase activity evidence_type: IEA original_reference_id: GO_REF:0000120 qualifier: enables supporting_entities: - ARBA:ARBA00027016 - InterPro:IPR000961 - EC:2.7.11.1 - PANTHER:PTN001122925 - UniRule:UR000416695 review: summary: >- Combined IEA (ARBA, InterPro AGC C-terminal domain, EC 2.7.11.1, PANTHER, UniRule). coWts is the Capsaspora Warts/LATS ortholog: it has a LATS-type MOB-binding region (CDD cd21774), a kinase domain with intact ATP-binding lysine and catalytic aspartate, and an AGC C-terminal extension. In animals Warts/LATS phosphorylates Yorkie/YAP/TAZ on HXRXXS motifs; coYki carries four such motifs, and the non-phosphorylatable coYki 4SA mutant is nuclear and phenocopies the coWts knockout [PMID:35659869, PMID:38517944]. action: ACCEPT reason: >- This is the core molecular activity of a Warts/LATS kinase and nothing in the sequence argues against it. The PANTHER component of this row comes from a node in PTHR22988 (Rho-effector kinase family) rather than the LATS clade, but serine/threonine kinase activity is shared by both families, so the misplacement does not affect this term. Note that no in vitro kinase assay or phosphorylation readout (phospho-specific antibody, Phos-tag) has been reported for coWts; the activity is inferred from domain conservation and the 4SA genetic mimicry, not measured. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:CDD"] supporting_text: CDD; cd21774; MobB_LATS; 1. - reference_id: PMID:35659869 supporting_text: >- In mammals and in Drosophila, YAP/TAZ/Yorkie is regulated by cytoplasmic sequestration in response to phosphorylation of HXRXXS motifs by the LATS/Warts kinases (Zheng and Pan, 2019). - reference_id: PMID:38517944 supporting_text: >- These results suggest that the phenotypes observed in coHpo and coWts mutants are due to coYki activation resulting from a loss of upstream kinase signaling. - term: id: GO:0005524 label: ATP binding evidence_type: IEA original_reference_id: GO_REF:0000120 qualifier: enables supporting_entities: - InterPro:IPR000719 - InterPro:IPR000961 - InterPro:IPR017441 - UniRule:UR001503341 review: summary: >- Domain-based assignment from the protein kinase domain and its ATP-binding signature (K740). action: ACCEPT reason: >- Kinase domain with conserved glycine-rich loop/ATP-binding lysine; ATP binding is required for the predicted catalytic activity. Accepted as a sound domain-level inference, although ATP binding has not been measured. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:InterPro"] supporting_text: InterPro; IPR017441; Protein_kinase_ATP_BS. - term: id: GO:0005737 label: cytoplasm evidence_type: IEA original_reference_id: GO_REF:0000120 qualifier: located_in supporting_entities: - ARBA:ARBA00026971 - UniProtKB-SubCell:SL-0086 - PANTHER:PTN001122925 review: summary: >- Combined ARBA/SubCell/PANTHER inference. The localization of coWts protein has not been examined. Its inferred substrate, coYki, is retained in the cytoplasm in wild-type cells and becomes nuclear when coWts is lost [PMID:38517944], consistent with coWts acting in the cytoplasm as Warts/LATS does in animals. action: ACCEPT reason: >- Cytoplasm is the expected compartment for a Warts/LATS kinase, which retains its substrate in the cytoplasm. The term is broad enough to be safe without direct localization data. Accepted as a homology-based location; it should be confirmed by tagging coWts. supported_by: - reference_id: PMID:38517944 supporting_text: >- Together, these results show that the Capsaspora Hippo kinase cascade regulates coYki by cytoplasmic sequestration, indicating a premetazoan origin of this regulatory activity of the Hippo pathway. - term: id: GO:0005856 label: cytoskeleton evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: located_in supporting_entities: - PANTHER:PTN001122925 review: summary: >- TreeGrafter transfer from PANTHER node PTN001122925. TreeGrafter places coWts in PTHR22988:SF71 "CITRON RHO-INTERACTING KINASE" (family PTHR22988, whose human members are ROCK1/2, MRCKs and CIT), not in the LATS clade where human LATS1/2 sit (PTHR24356). No localization of coWts has been reported. action: MARK_AS_OVER_ANNOTATED reason: >- The cytoskeletal location is characteristic of the Rho-effector kinases of PTHR22988, and reaches coWts because of the family placement rather than anything known about Warts/LATS. coWts's MOB-binding region and C-terminal kinase domain identify it as a LATS kinase. Animal LATS kinases are cytoplasmic with some cortical/centrosomal pools, so an association with cytoskeletal structures cannot be ruled out, but there is no evidence for it here. Not removed outright because the term is not demonstrably false. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR22988:SF71; CITRON RHO-INTERACTING KINASE; 1. - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:CDD"] supporting_text: CDD; cd21774; MobB_LATS; 1. propagation_review: root_cause: PROPAGATION_BAD failure_modes: - WRONG_ORTHOLOG_OR_PARALOG source_entities: - source_id: PANTHER:PTN001122925 source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET comment: >- TreeGrafter node in PTHR22988 (Rho-effector kinases); its cytoskeleton and actomyosin terms fit ROCK/MRCK/citron, while coWts is a Warts/LATS kinase whose human orthologs LATS1/2 are in PTHR24356 and receive IBAs from LATS node PTN002390470. - term: id: GO:0007010 label: cytoskeleton organization evidence_type: IEA original_reference_id: GO_REF:0000117 qualifier: involved_in supporting_entities: - ARBA:ARBA00026736 review: summary: >- ARBA machine-learning rule. In Capsaspora, coWts knockout cells show a subpopulation of elongated, contractile cells (rescued by a coWts transgene) and shorter filopodia within aggregates [PMID:38517944]. These effects are phenocopied by hyperactive coYki and require coYki transcriptional activity. action: MARK_AS_OVER_ANNOTATED reason: >- coWts influences cytoskeletal behaviour, but indirectly: the phenotypes are attributed to derepressed coYki-dependent transcription, not to coWts acting on cytoskeletal structures. By the participation test, coWts does not do the work of assembling or arranging the cytoskeleton. Process terms describing the signalling role (hippo signaling) capture its function better. The F-actin, blebbistatin and latrunculin analyses were also performed in the coHpo mutant rather than coWts. supported_by: - reference_id: PMID:38517944 supporting_text: >- This result indicates that these phenotypes are mediated by the transcriptional activity of coYki. - reference_id: PMID:38517944 supporting_text: >- As contractile cells in the coHpo mutant background tended to show a more elongated morphology than the coWts mutant, we focused on the coHpo mutant for further analysis. - term: id: GO:0016301 label: kinase activity evidence_type: IEA original_reference_id: GO_REF:0000104 qualifier: enables supporting_entities: - UniRule:UR000416695 review: summary: UniRule assignment of the general kinase activity parent term. action: KEEP_AS_NON_CORE reason: >- True but redundant with the more specific protein serine/threonine kinase activity annotation. Kept as a non-core parent. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS50011; PROTEIN_KINASE_DOM; 1. - term: id: GO:0016740 label: transferase activity evidence_type: IEA original_reference_id: GO_REF:0000104 qualifier: enables supporting_entities: - UniRule:UR000416695 review: summary: UniRule assignment of the very general transferase activity term. action: KEEP_AS_NON_CORE reason: >- True (a kinase transfers phosphate) but far too general to be informative. Kept as a non-core parent of protein serine/threonine kinase activity. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS50011; PROTEIN_KINASE_DOM; 1. - term: id: GO:0031032 label: actomyosin structure organization evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001122925 review: summary: >- TreeGrafter transfer from PANTHER node PTN001122925 in PTHR22988, the ROCK/MRCK/citron Rho-effector kinase family, whose members directly phosphorylate myosin light chain and regulators of actomyosin. coWts is a Warts/LATS kinase (MOB-binding region, C-terminal kinase domain) that falls into this family's HMM while its human orthologs LATS1/2 are classified in PTHR24356. The coWts knockout does show a myosin-sensitive contractile cell behaviour and denser aggregates [PMID:38517944], but actomyosin structures were imaged and perturbed only in coHpo mutants and wild type, and the phenotypes are attributed to coYki transcription. action: REMOVE reason: >- The term comes from a family-level placement with the Rho-effector kinases, which do organize actomyosin directly; it is not inherited from the Warts/LATS lineage. The Capsaspora data do not rescue it: coWts loss increases a contractile behaviour through coYki-dependent transcription (phenocopied by coYki 4SA, abolished by the TEAD-binding F123A mutation), there was no epistasis test, and the authors state that the role of myosin in these phenotypes remains unclear. coWts therefore acts upstream of, not in, actomyosin structure organization. This is the Track C pattern of a family-specific cytoskeletal process term transferred to a unicellular protein through a misassigned family. supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR22988; MYOTONIC DYSTROPHY S/T KINASE-RELATED; 1. - reference_id: PMID:38517944 supporting_text: >- This result indicates that these phenotypes are mediated by the transcriptional activity of coYki. - reference_id: PMID:38517944 supporting_text: >- Thus the specific role of myosin activity in the phenotypes observed in Hippo pathway mutants in Capsaspora remains unclear. propagation_review: root_cause: PROPAGATION_BAD failure_modes: - WRONG_ORTHOLOG_OR_PARALOG - ROLE_CONFLATION source_entities: - source_id: PANTHER:PTN001122925 source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET comment: >- PTHR22988 node (ROCK/MRCK/citron family); actomyosin organization is a direct activity of those Rho-effector kinases, but coWts belongs to the Warts/LATS clade, which acts on actomyosin only indirectly via Yorkie. - term: id: GO:0071944 label: cell periphery evidence_type: IEA original_reference_id: GO_REF:0000117 qualifier: located_in supporting_entities: - ARBA:ARBA00027443 review: summary: >- ARBA machine-learning rule. In animals Warts/LATS is recruited to the plasma membrane by Mats/MOB1 and Merlin/Kibra during activation; Capsaspora encodes Kibra and Merlin orthologs. The localization of coWts itself has not been examined. action: UNDECIDED reason: >- A membrane-proximal pool is plausible for a Warts/LATS kinase, but there are no Capsaspora data for or against it, and the rule-based assignment is not specific to the LATS clade. Neither acceptance nor removal is justified without localization of tagged coWts. supported_by: - reference_id: PMID:38517944 supporting_text: >- The Capsaspora genome encodes all components of the core Hippo pathway as well as some conserved upstream Hippo pathway regulators such as Kibra and NF2/Merlin - term: id: GO:1900181 label: negative regulation of protein localization to nucleus evidence_type: IEA original_reference_id: GO_REF:0000117 qualifier: involved_in supporting_entities: - ARBA:ARBA00092942 review: summary: >- ARBA rule. Directly supported in Capsaspora: in coWts knockout cells mScarlet-coYki is nuclear in most cells, whereas it is predominantly cytoplasmic in wild type; the effect is stronger than in coHpo knockouts [PMID:38517944]. action: ACCEPT reason: >- Restraining nuclear entry of Yorkie/YAP/TAZ is the canonical output of Warts/LATS and is shown genetically for coWts by the knockout (IMP-grade evidence from PMID:38517944, recorded here rather than as a duplicate row). Caveat: the mechanism (phosphorylation of coYki) is inferred from the 4SA mutant, not measured. supported_by: - reference_id: PMID:38517944 supporting_text: >- In contrast, the majority of coHpo-/- and coWts-/- cells showed nuclear levels of mScarlet-coYki greater than or equal to that seen in the cytoplasm (Figure 1C and D). - term: id: GO:0035329 label: hippo signaling evidence_type: IMP original_reference_id: PMID:38517944 review: summary: >- coWts is the Capsaspora Warts/LATS kinase of the Hippo cascade. Its knockout releases coYki to the nucleus, and its cytoskeletal and aggregate phenotypes are phenocopied by coYki 4SA (predicted Warts sites mutated) in a TEAD-binding-dependent manner, matching the canonical Hpo-Wts-Yki-Sd output. action: NEW reason: >- Participation test: Warts/LATS performs a step of the cascade (phosphorylation of Yorkie), so it is a participant rather than a regulated target. Comparator check: Drosophila wts and human LATS1/LATS2 carry GO:0035329 (experimental and IBA from LATS node PTN002390470). coWts received no hippo signaling IEA/IBA only because TreeGrafter placed it in PTHR22988 rather than the LATS clade. The IMP rests on the pathway readout (coYki nuclear localization) and the 4SA phenocopy. Limits: coHpo-to-coWts and coWts-to-coYki phosphorylation were not shown, and coWts may also have coHpo-independent activity. additional_reference_ids: - PMID:35659869 supported_by: - reference_id: PMID:38517944 supporting_text: >- Together, these results show that the Capsaspora Hippo kinase cascade regulates coYki by cytoplasmic sequestration, indicating a premetazoan origin of this regulatory activity of the Hippo pathway. - reference_id: PMID:38517944 supporting_text: >- These results suggest that the phenotypes observed in coHpo and coWts mutants are due to coYki activation resulting from a loss of upstream kinase signaling. - reference_id: PMID:35659869 supporting_text: >- In contrast, the majority of cells transfected with mScarlet-coYki 4SA showed uniform mScarlet signal throughout the cell or enriched mScarlet signal in the nucleus (Figure 5C and F). core_functions: - description: >- Warts/LATS-family serine/threonine kinase of the Capsaspora Hippo pathway that keeps the Yorkie/YAP/TAZ ortholog coYki in the cytoplasm, thereby restraining coYki-dependent transcription that otherwise induces a contractile elongated cell behaviour and denser cell packing within aggregates. Catalytic activity and coYki phosphorylation are inferred from domain conservation and the phenotype of a non-phosphorylatable coYki mutant; they have not been measured biochemically. molecular_function: id: GO:0004674 label: protein serine/threonine kinase activity directly_involved_in: - id: GO:0035329 label: hippo signaling - id: GO:1900181 label: negative regulation of protein localization to nucleus locations: - id: GO:0005737 label: cytoplasm supported_by: - reference_id: file:CAPO3/coWts/coWts-uniprot.txt record_section: ["DR:CDD"] supporting_text: CDD; cd21774; MobB_LATS; 1. - reference_id: PMID:38517944 supporting_text: >- In contrast, the majority of coHpo-/- and coWts-/- cells showed nuclear levels of mScarlet-coYki greater than or equal to that seen in the cytoplasm (Figure 1C and D). - reference_id: PMID:38517944 supporting_text: >- These results suggest that the phenotypes observed in coHpo and coWts mutants are due to coYki activation resulting from a loss of upstream kinase signaling. - reference_id: PMID:38517944 supporting_text: >- This result indicates that these phenotypes are mediated by the transcriptional activity of coYki. suggested_questions: - question: >- Does coWts phosphorylate coYki on its HXRXXS motifs in vitro and in cells, and does coHpo phosphorylate coWts on its activation-loop and hydrophobic-motif sites? - question: >- Why is the coYki nuclear-localization phenotype stronger in coWts than in coHpo mutants - which kinases (e.g. other STE20/MAP4K-like kinases) activate coWts independently of coHpo? - question: >- Where is coWts located in Capsaspora cells (cytoplasm, cortex, filopodia, adhesion foci), and is it recruited to membranes by Mats/MOB, Kibra or Merlin orthologs? - question: >- Should PANTHER place Warts/LATS orthologs such as coWts and Drosophila wts in PTHR24356 with human LATS1/2 rather than in PTHR22988 (Rho-effector kinases), so that they inherit the LATS-node IBAs (hippo signaling) instead of the ROCK/citron actomyosin terms? - question: >- Which coYki target genes (e.g. the actin-binding and laminin G-domain genes shared among the coHpo, coWts and coYki RNA-seq sets) mediate the contractile behaviour and cell-packing phenotype? - question: >- What explains the slower growth of coWts mutants in shaking culture - a coYki-independent function of coWts, or a viability effect of coYki hyperactivity? suggested_experiments: - hypothesis: coWts is an active kinase that phosphorylates coYki at its predicted Warts sites. description: >- In vitro kinase assay with immunopurified wild-type versus kinase-dead (K740A or D834A) coWts on recombinant coYki and coYki 4SA, plus Phos-tag or phospho-specific immunoblots of coYki in wild-type and coWts-/- Capsaspora cells. experiment_type: biochemical kinase assay - hypothesis: Kinase activity, not only the protein scaffold, is required for coWts function. description: >- Complement coWts-/- cells with wild-type versus kinase-dead coWts and score coYki nuclear localization, elongated-cell frequency and aggregate cell density. experiment_type: genetic complementation - hypothesis: coWts acts upstream of coYki for all scored phenotypes. description: >- Construct coWts coYki double mutants (once sequential marker recycling is available) or deplete coYki in coWts-/- cells and test whether contractile behaviour, cell packing and the filopodial defect are suppressed. experiment_type: genetic epistasis - hypothesis: coWts localizes to the cell cortex or adhesion foci. description: >- Express fluorescently tagged coWts from its endogenous locus or at low levels and image together with Lifeact and membrane (NMM) markers in adherent cells and aggregates. experiment_type: localization / imaging references: - id: GO_REF:0000002 title: Gene Ontology annotation through association of InterPro records with GO terms findings: [] - id: GO_REF:0000104 title: Electronic Gene Ontology annotations created by transferring manual GO annotations between related proteins based on shared sequence features findings: [] - id: GO_REF:0000117 title: Electronic Gene Ontology annotations created by ARBA machine learning models findings: [] - id: GO_REF:0000118 title: TreeGrafter-generated GO annotations findings: [] - id: GO_REF:0000120 title: Combined Automated Annotation using Multiple IEA Methods findings: [] - id: file:CAPO3/coWts/coWts-uniprot.txt title: UniProt record for coWts (A0A0D2VGR4) findings: - statement: >- coWts has a LATS-type MOB-binding region, a protein kinase domain with intact ATP-binding and catalytic residues, and an AGC C-terminal domain. record_section: ["DR:CDD"] supporting_text: CDD; cd21774; MobB_LATS; 1. - statement: >- PANTHER/TreeGrafter classifies coWts in the citron Rho-interacting kinase subfamily of the Rho-effector kinase family, not with human LATS1/2. record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR22988:SF71; CITRON RHO-INTERACTING KINASE; 1. - id: PMID:38517944 title: The Hippo kinase cascade regulates a contractile cell behavior and cell density in a close unicellular relative of animals. reference_review: relevance: HIGH correctness: VERIFIED review_notes: >- Primary paper for coWts; full text cached. Methods name the targeted gene as coWts (CAOG_00619); NCBI RefSeq XP_004365490.1 carries locus_tag CAOG_00619 and is cross-referenced by UniProt A0A0D2VGR4 (new tag CAOG_000619). Reports knockout, coYki localization, proliferation, cell morphology, aggregate and RNA-seq data. No biochemical kinase or phosphorylation measurements. findings: - statement: The coWts kinase domain was deleted from both alleles. supporting_text: >- For each gene, homologous recombination was used to replace the kinase domain of both WT alleles with selectable markers - statement: coWts loss increases nuclear coYki, more strongly than coHpo loss. supporting_text: >- Interestingly, we found that coWts-/- cells were significantly more likely to show nuclear mScarlet-coYki localization than coHpo-/- cells (Figure 1D) - statement: coWts is not a negative regulator of proliferation in Capsaspora. supporting_text: >- this result suggests that coWts is not a negative regulator of cell proliferation as in Drosophila and mammals, but instead is required for normal cell viability under some growth conditions - statement: >- coWts loss produces elongated contractile cells, rescued by a coWts transgene. supporting_text: >- The presence of elongated cells was rescued by transgenic expression of coHpo or coWts in the respective mutant (Figure 3A and B), demonstrating that the elongated cell phenotype was specifically due to the absence of these kinases. - statement: coWts mutant aggregates are less circular and more densely packed. supporting_text: >- Whereas the number of cells per unit area in coYki -/- aggregates was not significantly different from WT, the number of cells per area in coHpo -/- and coWts -/- was significantly increased relative to WT (Figure 5E). - statement: coWts mutant cells have shorter filopodia within aggregates. supporting_text: >- Whereas filopodia from coHpo -/- cells resembled those of WT and often extended more than a cell diameter distance within the aggregate, filopodia from coWts -/- cells were much shorter - statement: >- The phenotypes depend on coYki transcriptional activity; epistasis could not be tested. supporting_text: >- Since techniques are currently unavailable to create double mutants in Capsaspora, we could not test this possibility through classic genetic epistasis. - id: PMID:35659869 title: Genome editing in the unicellular holozoan Capsaspora owczarzaki suggests a premetazoan role for the Hippo pathway in multicellular morphogenesis. reference_review: relevance: MEDIUM correctness: VERIFIED review_notes: >- coYki knockout paper; establishes that coYki with its predicted Warts sites mutated (4SA) is nuclear, the basis for inferring coWts substrate and mechanism. coWts itself is not studied. findings: - statement: >- coYki with its four predicted Warts/LATS phosphorylation sites mutated is enriched in the nucleus. supporting_text: >- In contrast, the majority of cells transfected with mScarlet-coYki 4SA showed uniform mScarlet signal throughout the cell or enriched mScarlet signal in the nucleus (Figure 5C and F). - id: PMID:22832104 title: Premetazoan origin of the hippo signaling pathway. reference_review: relevance: LOW correctness: VERIFIED review_notes: >- Identifies the Capsaspora Hippo pathway components including a Wts homolog and tests Co-Yki, Co-Sd and Co-Hpo in Drosophila. Capsaspora Wts was aligned but not functionally tested; Co-Hpo phosphorylated Drosophila Wts. findings: - statement: Wts homologs are present across opisthokonts. supporting_text: >- Likewise, Wts homologues are present along all opisthokonts, except maybe for the ichthyosporeans, where the incompleteness of genome data makes it difficult to ensure its absence. - id: PMID:38729842 title: 'Assembling a Hippo: the evolutionary emergence of an animal developmental signaling pathway.' reference_review: relevance: LOW correctness: VERIFIED review_notes: Review; summarizes the coWts knockout phenotypes without new data. findings: - statement: >- Deleting coWts does not increase proliferation but increases an actomyosin-mediated contractile behaviour. supporting_text: >- Consistent with this result, targeted deletion of either Capsaspora Hippo (coHpo) or Capsaspora Warts (coWts) does not increase cell proliferation, but greatly increases the rate of occurrence of an actomyosin-mediated contractile behavior displayed by cells [66] (Figure 3A).