Transcription-independent RNA polymerase II dephosphorylation by the FCP1 carboxy-terminal domain phosphatase in Xenopus laevis early embryos.
The phosphorylation of the RNA polymerase II (RNAP II) carboxy-terminal domain (CTD) plays a key role in mRNA metabolism. The relative ratio of hyperphosphorylated RNAP II to hypophosphorylated RNAP II is determined by a dynamic equilibrium between CTD kinases and CTD phosphatase(s). The CTD is heavily phosphorylated in meiotic Xenopus laevis oocytes. In this report we show that the CTD undergoes fast and massive dephosphorylation upon fertilization. A cDNA was cloned and shown to code for a full-length xFCP1, the Xenopus orthologue of the FCP1 CTD phosphatases in humans and Saccharomyces cerevisiae. Two critical residues in the catalytic site were identified. CTD phosphatase activity was observed in extracts prepared from Xenopus eggs and cells and was shown to be entirely attributable to xFCP1. The CTD dephosphorylation triggered by fertilization was reproduced upon calcium activation of cytostatic factor-arrested egg extracts. Using immunodepleted extracts, we showed that this dephosphorylation is due to xFCP1. Although transcription does not occur at this stage, phosphorylation appears as a highly dynamic process involving the antagonist action of Xp42 mitogen-activated protein kinase and FCP1 phosphatase. This is the first report that free RNAP II is a substrate for FCP1 in vivo, independent from a transcription cycle.
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Génétique Moléculaire, UMR 8541 CNRS, Ecole Normale Supérieure, 75230 Paris Cedex 05, France.
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Dahmus
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Bensaude
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D000595:Amino Acid Sequence; D000818:Animals; D002457:Cell Extracts / Q000032:analysis; D003001:Cloning, Molecular; D004622:Embryo, Mammalian / Q000201:enzymology; D004625:Embryo, Nonmammalian; D019143:Evolution, Molecular; D006801:Humans; D008954:Models, Biological; D008969:Molecular Sequence Data; D010063:Ovum / Q000201:enzymology; D010710:Phosphates / Q000378:metabolism; D010749:Phosphoprotein Phosphatases / Q000235:genetics* / Q000502:physiology*; D010766:Phosphorylation; D012319:RNA Polymerase II / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D014158:Transcription, Genetic; D029867:Xenopus Proteins; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism*
D002457:Cell Extracts; D010710:Phosphates; D029867:Xenopus Proteins; D012319:RNA Polymerase II; C435123:CTD phosphatase protein, Xenopus; D010749:Phosphoprotein Phosphatases; C116022:carboxy-terminal domain phosphatase