title: Phospho-site mutants of the RNA Polymerase II C-terminal domain alter subtelomeric gene expression and chromatin modification state in fission yeast.
abstract: Eukaryotic gene expression requires that RNA Polymerase II (RNAP II) gain access to DNA in the context of chromatin. The C-terminal domain (CTD) of RNAP II recruits chromatin modifying enzymes to promoters, allowing for transcription initiation or repression. Specific CTD phosphorylation marks facilitate recruitment of chromatin modifiers, transcriptional regulators, and RNA processing factors during the transcription cycle. However, the readable code for recruiting such factors is still not fully defined and how CTD modifications affect related families of genes or regional gene expression is not well understood. Here, we examine the effects of manipulating the Y1S2P3T4S5P6S7heptapeptide repeat of the CTD of RNAP II in Schizosaccharomyces pombe by substituting non-phosphorylatable alanines for Ser2 and/or Ser7 and the phosphomimetic glutamic acid for Ser7. Global gene expression analyses were conducted using splicing-sensitive microarrays and validated via RT-qPCR. The CTD mutations did not affect pre-mRNA splicing or snRNA levels. Rather, the data revealed upregulation of subtelomeric genes and alteration of the repressive histone H3 lysine 9 methylation (H3K9me) landscape. The data further indicate that H3K9me and expression status are not fully correlated, suggestive of CTD-dependent subtelomeric repression mechansims that act independently of H3K9me levels.
authors.affiliation: Biology Department, Ithaca College, Ithaca, NY 14850, USA minada@ithaca.edu.
authors.forename: Maki
authors.initials: M
authors.lastname: Inada
authors.affiliation: Biology Department, Ithaca College, Ithaca, NY 14850, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Nichols
authors.affiliation: Department of Biochemistry and Biophysics, UCSF, San Francisco, CA 94158, USA.
authors.forename: Jahan-Yar
authors.initials: JY
authors.lastname: Parsa
authors.affiliation: Department of Biochemistry and Biophysics, UCSF, San Francisco, CA 94158, USA.
authors.forename: Christina M
authors.initials: CM
authors.lastname: Homer
authors.affiliation: Biology Department, Ithaca College, Ithaca, NY 14850, USA.
authors.forename: Ruby A
authors.initials: RA
authors.lastname: Benn
authors.affiliation: Biology Department, Ithaca College, Ithaca, NY 14850, USA.
authors.forename: Reyal S
authors.initials: RS
authors.lastname: Hoxie
authors.affiliation: Department of Biochemistry and Biophysics, UCSF, San Francisco, CA 94158, USA.
authors.forename: Hiten D
authors.initials: HD
authors.lastname: Madhani
authors.affiliation: Molecular Biology Program, Sloan-Kettering Institute, New York, NY 10065, USA.
authors.forename: Stewart
authors.initials: S
authors.lastname: Shuman
authors.affiliation: Department of Microbiology, Weill Cornell Medical College, New York, NY 10065, USA.
authors.forename: Beate
authors.initials: B
authors.lastname: Schwer
authors.affiliation: Department of Molecular Biology and Genetics, Cornell University, Ithaca, NY 14853, USA jpleiss@cornell.edu.
authors.forename: Jeffrey A
authors.initials: JA
authors.lastname: Pleiss
mesh_terms: D002843:Chromatin / Q000235:genetics* / Q000378:metabolism*; D016000:Cluster Analysis; D020869:Gene Expression Profiling; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006657:Histones; D008745:Methylation; D009154:Mutation; D010766:Phosphorylation; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D012319:RNA Polymerase II / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D012326:RNA Splicing; D012342:RNA, Small Nuclear / Q000378:metabolism; D015203:Reproducibility of Results; D012568:Schizosaccharomyces / Q000235:genetics / Q000378:metabolism; D017381:Spliceosomes / Q000378:metabolism
chemical_list: D002843:Chromatin; D006657:Histones; D012342:RNA, Small Nuclear; D012319:RNA Polymerase II