TAILIEUCHUNG - Báo cáo khoa học: Investigating RNA polymerase II carboxyl-terminal domain (CTD) phosphorylation ˆ Benoıt Palancade and Olivier Bensaude

Phosphorylation of RNA polymerase II’s largest subunit C-terminal domain (CTD) is a key event during mRNA metabolism. Numerous enzymes, including cell cycle-dependent kinases and TFIIF-dependent phosphatases tar-get the CTD. However, the repetitive nature of the CTD prevents determination of phosphorylated sites by conven-tional biochemistry methods. Fortunately, a panel of monoclonal antibodies is available that distinguishes between phosphorylated isoforms of RNA polymerase II’s (RNAP II) largest subunit | Eur. J. Biochem. 270 3859-3870 2003 FEBS 2003 doi REVIEW ARTICLE Investigating RNA polymerase II carboxyl-terminal domain CTD phosphorylation Benoit Palancade and Olivier Bensaude Génétique Moléculaire UMR 8541 CNRS Ecole Normale Superieure Paris France Phosphorylation of RNA polymerase II s largest subunit C-terminal domain CTD is a key event during mRNA metabolism. Numerous enzymes including cell cycledependent kinases and TFIIF-dependent phosphatases target the CTD. However the repetitive nature of the CTD prevents determination of phosphorylated sites by conventional biochemistry methods. Fortunately a panel of monoclonal antibodies is available that distinguishes between phosphorylated isoforms of RNA polymerase II s RNAP II largest subunit. Here we review how successful these tools have been in monitoring RNAP II phosphorylation changes in vivo by immunofluorescence chromatin immunoprecipitation and immunoblotting experiments. The CTD phosphorylation pattern is precisely modified as RNAP II progresses along the genes and is involved in sequential recruitment of RNA processing factors. One of the most popular anti-phosphoCTD Igs H5 has been proposed in several studies as a landmark of RNAP II molecules engaged in transcription. Finally we discuss how global RNAP II phosphorylation changes are affected by the physiological context such as cell stress and embryonic development. Keywords RNA polymerase II CTD-phosphorylation CTD-kinase CTD-phosphatase transcription mRNA processing. The C-terminal domain CTD of RNA polymerase II s RNAP II largest subunit is essential for transcription 1-3 for its function as enhancer 4 5 for organization of transcription foci within the nucleus 6 and for pre-mRNA processing 7 8 . The CTD consists of multiple repeats of a seven amino acid motif 9 10 Fig. 1 . This motif has been conserved during evolution in eukaryotes but the number of repeats varies depending on the species 26 in yeast 45 in flies

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