id: F2U943 gene_symbol: warts aliases: - PTSG_04961 - Warts - Wts - LATS homolog product_type: PROTEIN status: DRAFT taxon: id: NCBITaxon:946362 label: Salpingoeca rosetta description: >- warts (PTSG_04961) encodes the Warts/LATS-family (NDR subgroup, AGC group) serine/threonine protein kinase of the colonial choanoflagellate Salpingoeca rosetta. The 833-residue protein has a long, largely disordered N-terminal region, a C-terminal protein kinase domain (residues 456-763) with the conserved ATP-binding lysine and catalytic aspartate and an insert between the two kinase lobes, and an AGC-kinase C-terminal extension. It is the S. rosetta member of the choanoflagellate-plus-animal Warts clade that includes human LATS1/2 and Drosophila Warts. In animals Warts/LATS, activated by Hippo, phosphorylates the coactivator Yorkie/YAP on HXRXXS motifs and keeps it out of the nucleus; the S. rosetta Yorkie homolog carries four such motifs. In S. rosetta, warts knockout cells proliferate more slowly than wild type, the reverse of the animal overgrowth phenotype, yet form rosettes with about twice as many cells as wild type (up to about 60 versus at most 25) around a larger, more branched core of extracellular matrix. Warts and Yorkie both influence expression of extracellular matrix genes, including couscous. Its kinase activity and substrates have not been measured. existing_annotations: - term: id: GO:0000082 label: G1/S transition of mitotic cell cycle evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001220369 review: summary: >- TreeGrafter transfer. F2U943 was grafted onto PTN001220369, the Monosiga brevicollis Warts leaf, and inherits the IBD terms of its parent PTN002390470, a speciation node labelled "Metazoa-Choanoflagellida" whose descendants include human LATS1/2, Drosophila wts and Nematostella and M. brevicollis Warts (PANTHER 19 tree). The IBA for this term on human LATS1 cites mouse Lats2 and human LATS2. No cell-cycle phase analysis has been done in S. rosetta; warts knockout cells proliferate more slowly, the opposite of the animal loss-of-function phenotype [DOI:10.1101/2024.07.13.603360]. action: MARK_AS_OVER_ANNOTATED reason: >- G1/S transition is a eukaryote-wide process, so there is no taxon violation. But the donor evidence is a cell-cycle role of vertebrate LATS2 (mouse Lats2, human LATS2), which says little about a choanoflagellate. In S. rosetta, loss of warts slows rather than accelerates proliferation, and no cell-cycle phase was measured, so nothing supports a direct role in the G1/S transition. Kept as an over-annotation rather than removed, because the IBD placement at the Choanozoa node is a phylogenetic judgement I cannot refute with S. rosetta data. supported_by: - reference_id: DOI:10.1101/2024.07.13.603360 supporting_text: >- On the other hand, hippopac1 and warts pac1 KO clones proliferated markedly slower - reference_id: DOI:10.1101/2024.07.13.603360 supporting_text: >- down proliferation in S. rosetta, which is the opposite of their animal loss-of-function phenotype propagation_review: root_cause: PROPAGATION_BAD failure_modes: - CONTEXT_OR_TISSUE_MISMATCH source_entities: - source_id: PANTHER:PTN002390470 source_label: Warts/LATS, Metazoa-Choanoflagellida node source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET comment: >- IBD supported by vertebrate LATS2 cell-cycle evidence (MGI Lats2, human LATS2); sound for vertebrates, but no evidence the role extends to choanoflagellates. - source_id: PANTHER:PTN001220369 source_label: M. brevicollis Warts leaf source_status: NOT_RELEVANT comment: >- Graft point only; carries no experimental annotation of its own. - 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 assignment from the protein kinase ATP-binding signature (ATP-binding residue K485). action: KEEP_AS_NON_CORE reason: >- True but uninformative parent of ATP binding, itself a component of the kinase activity. No binding measurements exist. supported_by: - reference_id: file:SALRS/warts/warts-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 aspartate (D579, 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 better describes a Warts/LATS AGC kinase. Kept as a true, less specific parent. supported_by: - reference_id: file:SALRS/warts/warts-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: - InterPro:IPR000961 - EC:2.7.11.1 - PANTHER:PTN001220369 - UniRule:UR000416695 review: summary: >- Combined IEA (InterPro AGC C-terminal domain, EC 2.7.11.1, PANTHER, UniRule). F2U943 is classified in PANTHER PTHR24356:SF418 "SERINE_THREONINE-PROTEIN KINASE WARTS", the subfamily of human LATS1/2, and has an intact kinase domain (ATP-binding K485, catalytic D579) followed by an AGC C-terminal domain. In animals Warts/LATS phosphorylates Yorkie/YAP on HXRXXS motifs [PMID:35659869]. action: ACCEPT reason: >- This is the core molecular activity of a Warts/LATS kinase, and the catalytic residues are retained. Unlike Capsaspora coWts, whose TreeGrafter placement is in the Rho-effector kinase family, the S. rosetta protein sits in the LATS clade, so the family-level inference is sound. Kinase activity has not been assayed for the S. rosetta protein. supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR24356:SF418; SERINE_THREONINE-PROTEIN KINASE WARTS; 1. - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS00108; PROTEIN_KINASE_ST; 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). - 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 (K485). action: ACCEPT reason: >- Kinase domain with the conserved ATP-binding lysine; ATP binding is required for the predicted catalytic activity. Sound domain-level inference, not measured. supported_by: - reference_id: file:SALRS/warts/warts-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 review: summary: >- ARBA rule and UniProt subcellular-location keyword. Warts/LATS kinases act in the cytoplasm, where they retain phosphorylated Yorkie/YAP. No localization data exist for the S. rosetta protein. action: ACCEPT reason: >- Cytoplasm is the expected compartment for a Warts/LATS kinase, and the term is broad enough to be safe without direct data. Accepted as a homology-based location. supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["CC:SUBCELLULAR LOCATION"] supporting_text: 'SUBCELLULAR LOCATION: Cytoplasm' - 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). - 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. supported_by: - reference_id: file:SALRS/warts/warts-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 too general to be informative. Kept as a non-core parent of protein serine/threonine kinase activity. supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["DR:PROSITE"] supporting_text: PROSITE; PS50011; PROTEIN_KINASE_DOM; 1. - term: id: GO:0035329 label: hippo signaling evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001220369 review: summary: >- TreeGrafter transfer of the IBD placed at PTN002390470 (Warts/LATS, "Metazoa-Choanoflagellida"), supported by experimental annotations on Drosophila wts, human LATS1/2 and mouse Lats. In S. rosetta, Warts and Yorkie regulate several extracellular matrix genes [PMID:41037400], the Yorkie homolog carries four HXRXXS Warts motifs [PMID:38729842], and in Capsaspora the Warts homolog restrains nuclear entry of the Yorkie homolog [PMID:38517944]. action: ACCEPT reason: >- Warts/LATS performs a step of the hippo signaling cascade (phosphorylating Yorkie), and the IBD node includes choanoflagellates. That placement is consistent with Capsaspora evidence that the Hippo-Warts-Yorkie cascade predates animals, so the family-level inference is sound and the term describes the core role. Caveats: no S. rosetta experiment shows Warts phosphorylating Yorkie or controlling its localization; hippo knockout does not phenocopy the warts rosette-size phenotype; and the predicted TEAD-binding region of S. rosetta Yorkie is poorly conserved. A NEW IMP is not added: the knockout phenotypes (rosette size, proliferation) are downstream outcomes and do not test participation in the cascade itself. supported_by: - reference_id: PMID:41037400 supporting_text: >- RNA sequencing revealed that Warts and Yorkie regulated several extracellular matrix genes involved in multicellularity (including couscous) - reference_id: PMID:38729842 supporting_text: >- srYki has 4 putative Warts phosphorylation sites - 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. propagation_review: root_cause: NO_FAILURE_CORE source_entities: - source_id: PANTHER:PTN002390470 source_label: Warts/LATS, Metazoa-Choanoflagellida node source_status: SUPPORTS_TRANSFER comment: >- IBD supported by fly wts and mammalian LATS1/2; the cascade is shown to predate animals in Capsaspora, so inheritance at the Choanozoa node is plausible. - term: id: GO:0035556 label: intracellular signal transduction evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001220369 review: summary: >- TreeGrafter transfer of a general signalling term from the PTHR24356 tree; parent of hippo signaling. action: KEEP_AS_NON_CORE reason: >- True for a signalling kinase but less informative than hippo signaling, which is also annotated. Kept as a non-core parent. supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR24356:SF418; SERINE_THREONINE-PROTEIN KINASE WARTS; 1. - term: id: GO:0043065 label: positive regulation of apoptotic process evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001220369 review: summary: >- TreeGrafter transfer of an IBD at PTN002390470 whose experimental support is Drosophila wts (FBgn0011739), where Warts promotes apoptosis by inactivating Yorkie and so lowering expression of the caspase inhibitor diap1. No cell death has been examined in S. rosetta warts knockouts; the enlarged rosettes were described with a slower, not faster, proliferation rate [DOI:10.1101/2024.07.13.603360]. action: MARK_AS_OVER_ANNOTATED reason: >- The term is permitted for opisthokonts, so it is not a taxon violation, and a role in cell survival in a choanoflagellate is not impossible. But the donor biology is Yorkie-dependent control of an animal anti-apoptotic programme in growing tissue. There is no evidence that choanoflagellate Warts or Yorkie regulate apoptosis, and the Yorkie homolog's TEAD-binding region is poorly conserved, so transfer of this tissue-growth-control output is unsupported. Not removed, because no data contradict it. supported_by: - reference_id: PMID:38729842 supporting_text: >- The degree to which choanoflagellate Yorkie orthologs interact with Sd/TEAD proteins is therefore currently unclear - reference_id: DOI:10.1101/2024.07.13.603360 supporting_text: >- down proliferation in S. rosetta, which is the opposite of their animal loss-of-function phenotype propagation_review: root_cause: PROPAGATION_BAD failure_modes: - CONTEXT_OR_TISSUE_MISMATCH source_entities: - source_id: PANTHER:PTN002390470 source_label: Warts/LATS, Metazoa-Choanoflagellida node source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET comment: >- Seeded by Drosophila wts tissue-growth and apoptosis phenotypes; no choanoflagellate data on apoptosis. - term: id: GO:0046620 label: regulation of organ growth evidence_type: IEA original_reference_id: GO_REF:0000118 qualifier: involved_in supporting_entities: - PANTHER:PTN001220369 review: summary: >- TreeGrafter transfer of an IBD at PTN002390470 seeded by mouse Lats2 (MGI:1354386). GO defines the term as modulating "growth of an organ of an organism"; it is a child of regulation of developmental growth and regulation of multicellular organismal process. S. rosetta is a choanoflagellate that lives as single cells or as clonal rosettes of a few dozen cells; it has no organs. Its warts knockout does enlarge rosettes [DOI:10.1101/2024.07.13.603360], but a rosette is not an organ. action: REMOVE reason: >- An organ is a multicellular anatomical structure of an animal or plant body, which choanoflagellates do not have, so the term cannot be true of this protein whatever its function. The source annotation is sound for mice. The fault is that an organ-level outcome term was placed at a node ("Metazoa-Choanoflagellida") that includes unicellular organisms, while the GO term's taxon constraints exclude only bacteria, archaea and two yeasts. The rosette-size phenotype is the closest S. rosetta counterpart, but no GO term fits it (there is no rosette term; see the NTR drafted in the rosetteless review). It is also a downstream outcome of altered extracellular matrix gene expression, not work done by Warts, so no replacement process term is proposed. supported_by: - reference_id: DOI:10.1101/2024.07.13.603360 supporting_text: >- warts pac1 cells grew into giant rosettes containing about twice as many cells as wild-type ones (21.1 ± 4.4 versus 10.9 ± 3.8 cells per rosette, respectively) - reference_id: PMID:41037400 supporting_text: >- suggesting that Hippo signaling regulates multicellular size in choanoflagellates by modulating matrix secretion propagation_review: root_cause: PROPAGATION_BAD failure_modes: - LINEAGE_OR_TAXON_MISMATCH source_entities: - source_id: PANTHER:PTN002390470 source_label: Warts/LATS, Metazoa-Choanoflagellida node source_status: SUPPORTS_SOURCE_BUT_NOT_TARGET comment: >- Organ-growth IBD seeded by mouse Lats2; valid for animals, but the node spans choanoflagellates, which have no organs. - source_id: PANTHER:PTN001220369 source_label: M. brevicollis Warts leaf source_status: NOT_RELEVANT comment: Graft point only; M. brevicollis also receives this IBA and has no organs either. - 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 upstream adaptors during activation. The localization of S. rosetta Warts has not been examined. action: UNDECIDED reason: >- A membrane-proximal pool is plausible for a Warts/LATS kinase, but there are no S. rosetta data for or against it, and the rule is not specific to the LATS clade. supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["CC:SUBCELLULAR LOCATION"] supporting_text: 'SUBCELLULAR LOCATION: Cytoplasm' core_functions: - description: >- Warts/LATS-family serine/threonine protein kinase of the S. rosetta Hippo pathway, inferred from its LATS-clade family placement to restrain the Yorkie homolog, which carries four Warts-consensus HXRXXS motifs. In S. rosetta, Warts and Yorkie both influence extracellular matrix gene expression, including couscous. Loss of Warts roughly doubles the number of cells per rosette, with an enlarged and more branched matrix core, and slows cell proliferation. Kinase activity, Yorkie phosphorylation and activation by Hippo have not been measured. molecular_function: id: GO:0004674 label: protein serine/threonine kinase activity directly_involved_in: - id: GO:0035329 label: hippo signaling locations: - id: GO:0005737 label: cytoplasm supported_by: - reference_id: file:SALRS/warts/warts-uniprot.txt record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR24356:SF418; SERINE_THREONINE-PROTEIN KINASE WARTS; 1. - reference_id: PMID:41037400 supporting_text: >- RNA sequencing revealed that Warts and Yorkie regulated several extracellular matrix genes involved in multicellularity (including couscous) - reference_id: DOI:10.1101/2024.07.13.603360 supporting_text: >- that ECM core appeared larger and with a more conspicuously branched geometry in warts pac1 suggested_questions: - question: >- Does S. rosetta Warts phosphorylate the Yorkie homolog on its four HXRXXS motifs, and is Yorkie more nuclear in warts knockout cells? - question: >- Are the enlarged rosettes of warts knockouts caused by Yorkie? yorkie knockout rosettes are of normal size, so a warts yorkie double knockout is the key epistasis test. - question: >- Why does hippo knockout not phenocopy the warts rosette-size phenotype? Which kinases activate Warts in S. rosetta? - question: >- Do larger warts knockout rosettes arise from more cell divisions before rosette release, reduced cell loss, or altered matrix secretion (e.g. via couscous)? - question: >- Should GO add taxon constraints restricting organ-level growth terms such as regulation of organ growth (GO:0046620) to taxa with organs, or should PAINT place the organ-growth IBD at a metazoan rather than a Metazoa-plus-choanoflagellate node? suggested_experiments: - hypothesis: S. rosetta Warts is an active kinase that phosphorylates Yorkie at HXRXXS motifs. description: >- In vitro kinase assay of immunopurified wild-type versus kinase-dead (K485A or D579A) Warts on recombinant Yorkie and a four-site S-to-A Yorkie mutant, plus Phos-tag immunoblots of Yorkie in wild-type and warts knockout cells. experiment_type: biochemical kinase assay - hypothesis: The warts rosette-size phenotype is mediated by Yorkie. description: >- Generate warts yorkie double knockouts with the selection-based cassette method and count cells per rosette; express a non-phosphorylatable Yorkie in wild-type cells to test whether it phenocopies warts loss. experiment_type: genetic epistasis - hypothesis: Warts kinase activity is required to limit rosette size. description: >- Complement warts knockout cells with wild-type or kinase-dead Warts and score cells per rosette, ECM core morphology and doubling time. experiment_type: genetic complementation 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:SALRS/warts/warts-uniprot.txt title: UniProt record for warts (F2U943) findings: - statement: >- PANTHER classifies F2U943 in the Warts subfamily of PTHR24356, with human LATS1/2. record_section: ["DR:PANTHER"] supporting_text: PANTHER; PTHR24356:SF418; SERINE_THREONINE-PROTEIN KINASE WARTS; 1. - id: DOI:10.1101/2024.07.13.603360 title: A fast and robust gene knockout method for Salpingoeca rosetta informs the genetics of choanoflagellate multicellular development reference_review: relevance: HIGH correctness: VERIFIED review_notes: >- bioRxiv preprint (July 2024), full text cached from the PDF. Reports CRISPR knockout of the S. rosetta Hippo, Warts and Yorkie homologs (two clones each), growth curves, rosette-size counts and ECM imaging of warts knockout rosettes. The cached version does not give locus IDs; PTSG_04961 as the warts locus was supplied from the preprint's gRNA-design sentence, which is not in this cached version. F2U943 is the only S. rosetta protein in the PANTHER Warts subfamily, which supports the mapping. Published as PMID:41037400. findings: - statement: warts knockout rosettes contain about twice as many cells as wild type. supporting_text: >- warts pac1 cells grew into giant rosettes containing about twice as many cells as wild-type ones (21.1 ± 4.4 versus 10.9 ± 3.8 cells per rosette, respectively) - statement: warts knockout rosettes reach up to about 60 cells. supporting_text: >- as many as 60 cells, while their wild type counterparts did not exceed 25 cells - statement: warts knockout rosettes have a larger, more branched ECM core. supporting_text: >- that ECM core appeared larger and with a more conspicuously branched geometry in warts pac1 - statement: warts knockout cells proliferate more slowly than wild type. supporting_text: >- On the other hand, hippopac1 and warts pac1 KO clones proliferated markedly slower - statement: All knockout clones still develop into rosettes. supporting_text: >- all six KO clones reliably developed into rosettes, as did wild-type cells - statement: The authors identify warts as a genetic regulator of rosette size. supporting_text: >- These observations pinpoint warts as a genetic regulator of S. rosetta rosette size - id: PMID:41037400 title: A selection-based knockout approach for a choanoflagellate reveals regulation of multicellular development by Hippo signaling. full_text_unavailable: true reference_review: relevance: HIGH correctness: VERIFIED review_notes: >- Published version of the preprint; abstract only cached. Adds RNA-seq showing that Warts and Yorkie regulate extracellular matrix genes including couscous. findings: - statement: warts knockout rosettes are larger than wild type. supporting_text: >- Interestingly, warts-KO rosettes were consistently larger than their wild-type counterparts. - statement: Warts and Yorkie regulate extracellular matrix genes, including couscous. supporting_text: >- RNA sequencing revealed that Warts and Yorkie regulated several extracellular matrix genes involved in multicellularity (including couscous) - 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: LOW correctness: VERIFIED review_notes: Background on the Warts/LATS mechanism (HXRXXS phosphorylation of Yorkie/YAP). findings: - statement: Warts/LATS phosphorylates Yorkie/YAP/TAZ on HXRXXS motifs. 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). - 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: MEDIUM correctness: VERIFIED review_notes: >- Capsaspora coWts knockout; shows a premetazoan Warts restraining Yorkie localization. Not about S. rosetta. findings: - statement: The Capsaspora Hippo kinase cascade keeps the Yorkie homolog cytoplasmic. 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. - id: PMID:22832104 title: Premetazoan origin of the hippo signaling pathway. reference_review: relevance: LOW correctness: VERIFIED review_notes: Comparative genomics; Wts homologs present across opisthokonts. 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: MEDIUM correctness: VERIFIED review_notes: >- Review; describes the S. rosetta Yorkie homolog (four putative Warts sites, poorly conserved TEAD-binding region), relevant to the predicted Warts substrate. findings: - statement: S. rosetta Yorkie has four putative Warts phosphorylation sites. supporting_text: srYki has 4 putative Warts phosphorylation sites - statement: It is unclear whether choanoflagellate Yorkie homologs bind Sd/TEAD. supporting_text: >- The degree to which choanoflagellate Yorkie orthologs interact with Sd/TEAD proteins is therefore currently unclear