title: CDK5 is present in sea urchin and starfish eggs and embryos and can interact with p35, cyclin E and cyclin B3.
abstract: While most cyclin-dependent kinases (CDKs) are involved in cell cycle control, CDK5 is mostly known for crucial functions in neurogenesis. However, we cloned sea urchin CDK5 from a two-cell stage cDNA library and found that the protein is present in eggs and embryos, up to the pluteus stage, but without associated kinase activity. To investigate the potential for nonneuronal roles, we screened a starfish cDNA library with the yeast two-hybrid system, for possible CDK5 partners. Interactions with clones expressing part of cyclin B3 and cyclin E proteins were found and the full-length cyclins were cloned. These interactions were verified in vitro but not in extracts of starfish oocytes and embryos, at any stages, despite the presence of detectable amounts of CDK5, cyclin B3, and cyclin E. We then looked for p35, the CDK5-specific activator, and cloned the sea urchin ortholog. A sea urchin-specific anomaly in the amino acid sequence is the absence of N-terminal myristoylation signal, but nucleotide environment analysis suggests a much higher probability of translation initiation on the second methionine(Met44), that is associated with a conserved myristoylation signal. p35 was found to associate with CDK5 and, when bacterially produced, to confer protein kinase activity to CDK5 immunoprecipitated from sea urchin eggs and embryos. However, p35 mRNA expression was found to begin only at the end of the blastula stage, and the protein was undetectable at any embryonic stage, suggesting a neuronal role beginning in late larval stages.
authors.affiliation: Laboratoire Arago, Université Pierre et Marie Curie-Paris6, Banyuls-sur-Mer, France.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Lozano
authors.forename: Philippe
authors.initials: P
authors.lastname: Schatt
authors.forename: Valérie
authors.initials: V
authors.lastname: Vergé
authors.forename: Jérôme
authors.initials: J
authors.lastname: Gobinet
authors.forename: Vincent
authors.initials: V
authors.lastname: Villey
authors.forename: Gérard
authors.initials: G
authors.lastname: Peaucellier
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D019926:Cyclin B / Q000378:metabolism*; D019927:Cyclin E / Q000378:metabolism*; D051360:Cyclin-Dependent Kinase 5 / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004531:Eggs / Q000032:analysis; D004625:Embryo, Nonmammalian / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D010770:Phosphotransferases / Q000378:metabolism*; D012333:RNA, Messenger / Q000032:analysis; D012617:Sea Urchins / Q000737:chemistry / Q000196:embryology / Q000378:metabolism*; D016415:Sequence Alignment; D013215:Starfish / Q000737:chemistry* / Q000196:embryology
chemical_list: D019926:Cyclin B; D019927:Cyclin E; D012333:RNA, Messenger; D010770:Phosphotransferases; D051360:Cyclin-Dependent Kinase 5