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Incomplete RNA polymerase II phosphorylation in Xenopus laevis early embryos

Benoit Palancade¹, Sylvain Bellier^1,*, Geneviève Almouzni² and Olivier Bensaude^1,

¹ Génétique Moléculaire, UMR 8541 CNRS, Ecole Normale Supérieure, 46 rue d’Ulm, 75230 Paris Cedex 05, France
² Dynamique de la Chromatine, UMR 144 CNRS, Institut Curie, 26 rue d’Ulm, 75231 Paris Cedex 05, France
^* Present address: Service de Biochimie, Ecole Nationale Vétérinaire, 7 Av. du Général De Gaulle, 94704 Maisons-Alfort Cedex, France

Author for correspondence (e-mail: bensaude{at}biologie.ens.fr)

Accepted April 5, 2001

Phosphorylation of RNA polymerase II largest subunit on its C-terminal domain (CTD) heptapeptide repeats has been shown to play a key role in the regulation of mRNA synthesis and processing. In many higher metazoans, early embryos do not synthesise mRNAs during the first cell cycles following fertilisation. Transcription resumes and becomes an absolute requirement for development after several cell cycles characteristic of each species. Therefore, CTD phosphorylation has been investigated during early development of the African clawed-frog Xenopus laevis. Fertilisation is shown to trigger an abrupt dephosphorylation of the CTD. Phosphorylation of the CTD resumes concurrently with the mid-blastula transition (MBT). Both are advanced with polyspermy and increased temperatures; they do not occur when replication is impaired with aphidicolin. In Xenopus laevis somatic cells, a set of monoclonal antibodies defined distinct phosphoepitopes on the CTD. Two of them were absent before the MBT indicating that the CTD lacks the phosphorylation at the serine-2 position of the heptapeptide. The possible contribution of RNA polymerase II phosphorylation to the developmental-regulation of maternal mRNA processing in embryos is discussed.

Key words: RNA polymerase, phosphorylation, mid-blastula transition, zygotic gene activation.

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