title: Distinct functions of maternal and somatic Pat1 protein paralogs.
abstract: We previously identified Xenopus Pat1a (P100) as a member of the maternal CPEB RNP complex, whose components resemble those of P-(rocessing) bodies, and which is implicated in translational control in Xenopus oocytes. Database searches have identified Pat1a proteins in other vertebrates, as well as paralogous Pat1b proteins. Here we characterize Pat1 proteins, which have no readily discernable sequence features, in Xenopus oocytes, eggs, and early embryos and in human tissue culture cells. xPat1a and 1b have essentially mutually exclusive expression patterns in oogenesis and embryogenesis. xPat1a is degraded during meiotic maturation, via PEST-like regions, while xPat1b mRNA is translationally activated at GVBD by cytoplasmic polyadenylation. Pat1 proteins bind RNA in vitro, via a central domain, with a preference for G-rich sequences, including the NRAS 5' UTR G-quadruplex-forming sequence. When tethered to reporter mRNA, both Pat proteins repress translation in oocytes. Indeed, both epitope-tagged proteins interact with the same components of the CPEB RNP complex, including CPEB, Xp54, eIF4E1b, Rap55B, and ePAB. However, examining endogenous protein interactions, we find that in oocytes only xPat1a is a bona fide component of the CPEB RNP, and that xPat1b resides in a separate large complex. In tissue culture cells, hPat1b localizes to P-bodies, while mPat1a-GFP is either found weakly in P-bodies or disperses P-bodies in a dominant-negative fashion. Altogether we conclude that Pat1a and Pat1b proteins have distinct functions, mediated in separate complexes. Pat1a is a translational repressor in oocytes in a CPEB-containing complex, and Pat1b is a component of P-bodies in somatic cells.
authors.affiliation: Department of Biochemistry, University of Cambridge, Cambridge CB21QW, United Kingdom.
authors.forename: Aline
authors.initials: A
authors.lastname: Marnef
authors.forename: Maria
authors.initials: M
authors.lastname: Maldonado
authors.forename: Anthony
authors.initials: A
authors.lastname: Bugaut
authors.forename: Shankar
authors.initials: S
authors.lastname: Balasubramanian
authors.forename: Michel
authors.initials: M
authors.lastname: Kress
authors.forename: Dominique
authors.initials: D
authors.lastname: Weil
authors.forename: Nancy
authors.initials: N
authors.lastname: Standart
mesh_terms: D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D005260:Female; D006801:Humans; D051379:Mice; D009865:Oocytes / Q000378:metabolism*; D010802:Phylogeny; D011485:Protein Binding; D014176:Protein Biosynthesis; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger; D016601:RNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D029867:Xenopus Proteins / Q000235:genetics / Q000378:metabolism*; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism*
chemical_list: D004268:DNA-Binding Proteins; C553601:PATL1 protein, Xenopus; C526510:PATL1 protein, human; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D029867:Xenopus Proteins; C075198:PATL2 protein, Xenopus; D012313:RNA
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