Tyrosine phosphorylation of RNA polymerase II CTD is associated with antisense promoter transcription and active enhancers in mammalian cells.
In mammals, the carboxy-terminal domain (CTD) of RNA polymerase (Pol) II consists of 52 conserved heptapeptide repeats containing the consensus sequence Tyr1-Ser2-Pro3-Thr4-Ser5-Pro6-Ser7. Post-translational modifications of the CTD coordinate the transcription cycle and various steps of mRNA maturation. Here we describe Tyr1 phosphorylation (Tyr1P) as a hallmark of promoter (5' associated) Pol II in mammalian cells, in contrast to what was described in yeast. Tyr1P is predominantly found in antisense orientation at promoters but is also specifically enriched at active enhancers. Mutation of Tyr1 to phenylalanine (Y1F) prevents the formation of the hyper-phosphorylated Pol IIO form, induces degradation of Pol II to the truncated Pol IIB form, and results in a lethal phenotype. Our results suggest that Tyr1P has evolved specialized and essential functions in higher eukaryotes associated with antisense promoter and enhancer transcription, and Pol II stability.DOI: http://dx.doi.org/10.7554/eLife.02105.001.
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France Institut de Génétique Moléculaire de Montpellier (IGMM), CNRS-UMR5535, Montpellier, France.
Nicolas
N
Descostes
Department of Molecular Epigenetics, Helmholtz Center Munich, Center of Integrated Protein Science Munich, Munich, Germany.
Martin
M
Heidemann
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France.
Lionel
L
Spinelli
Department of Molecular Epigenetics, Helmholtz Center Munich, Center of Integrated Protein Science Munich, Munich, Germany.
Roland
R
Schüller
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France Institut de Génétique Moléculaire de Montpellier (IGMM), CNRS-UMR5535, Montpellier, France.
Muhammad Ahmad
MA
Maqbool
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France.
Romain
R
Fenouil
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France.
Frederic
F
Koch
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France.
Charlène
C
Innocenti
Centre Nacional D'Anàlisi Genòmica, Barcelona, Spain.
Marta
M
Gut
Centre Nacional D'Anàlisi Genòmica, Barcelona, Spain.
Ivo
I
Gut
Department of Molecular Epigenetics, Helmholtz Center Munich, Center of Integrated Protein Science Munich, Munich, Germany eick@helmholtz-muenchen.de.
Dirk
D
Eick
Centre d'Immunologie de Marseille-Luminy, Université Aix-Marseille, Marseille, France Centre National de la Recherche Scientifique (CNRS) UMR6102, Marseille, France Inserm U631, Marseille, France Institut de Génétique Moléculaire de Montpellier (IGMM), CNRS-UMR5535, Montpellier, France jean-christophe.andrau@igmm.cnrs.fr.
Jean-Christophe
JC
Andrau
D016375:Antisense Elements (Genetics); D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004742:Enhancer Elements, Genetic; D006801:Humans; D009154:Mutation; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D012319:RNA Polymerase II / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014443:Tyrosine / Q000378:metabolism*
D016375:Antisense Elements (Genetics); D014443:Tyrosine; D012319:RNA Polymerase II
RNA polymerase II; antisense transcription; carboxyl terminal domain; enhancers; initiation; transcription