title: Function and control of RNA polymerase II C-terminal domain phosphorylation in vertebrate transcription and RNA processing.
abstract: The C-terminal domain of the RNA polymerase II largest subunit (the Rpb1 CTD) is composed of tandem heptad repeats of the consensus sequence Y(1)S(2)P(3)T(4)S(5)P(6)S(7). We reported previously that Thr 4 is phosphorylated and functions in histone mRNA 3'-end formation in chicken DT40 cells. Here, we have extended our studies on Thr 4 and to other CTD mutations by using these cells. We found that an Rpb1 derivative containing only the N-terminal half of the CTD, as well as a similar derivative containing all-consensus repeats (26r), conferred full viability, while the C-terminal half, with more-divergent repeats, did not, reflecting a strong and specific defect in snRNA 3'-end formation. Mutation in 26r of all Ser 2 (S2A) or Ser 5 (S5A) residues resulted in lethality, while Ser 7 (S7A) mutants were fully viable. While S2A and S5A cells displayed defects in transcription and RNA processing, S7A cells behaved identically to 26r cells in all respects. Finally, we found that Thr 4 was phosphorylated by cyclin-dependent kinase 9 in cells and dephosphorylated both in vitro and in vivo by the phosphatase Fcp1.
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York, USA.
authors.forename: Jing-Ping
authors.initials: JP
authors.lastname: Hsin
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York, USA.
authors.forename: Kehui
authors.initials: K
authors.lastname: Xiang
authors.affiliation: Department of Biological Sciences, Columbia University, New York, New York, USA jlm2@columbia.edu.
authors.forename: James L
authors.initials: JL
authors.lastname: Manley
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002645:Chickens; D042863:Cyclin-Dependent Kinase 9 / Q000235:genetics*; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D010749:Phosphoprotein Phosphatases / Q000235:genetics*; D010766:Phosphorylation / Q000235:genetics; D021122:Protein Subunits / Q000235:genetics; D012319:RNA Polymerase II / Q000235:genetics*; D012323:RNA Processing, Post-Transcriptional; D012342:RNA, Small Nuclear / Q000096:biosynthesis / Q000235:genetics; D013912:Threonine / Q000737:chemistry* / Q000235:genetics; D014158:Transcription, Genetic
chemical_list: D021122:Protein Subunits; D012342:RNA, Small Nuclear; D013912:Threonine; C467969:CDK9 protein, human; D042863:Cyclin-Dependent Kinase 9; D012319:RNA Polymerase II; C410949:RNA polymerase II largest subunit; C000588909:Fcp1 protein, human; D010749:Phosphoprotein Phosphatases
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