Sus1 is recruited to coding regions and functions during transcription elongation in association with SAGA and TREX2.
Gene transcription, RNA biogenesis, and mRNA transport constitute a complicated process essential for all eukaryotic cells. The transcription/export factor Sus1 plays a key role in coupling transcription activation with mRNA export, and it resides in both the SAGA and TREX2 complexes. Moreover, Sus1 is responsible for GAL1 gene gating at ... the nuclear periphery, which is important for its transcriptional status. Here, we show that Sus1 is required during transcription elongation and is associated with the elongating form of RNA Polymerase II (RNAP II) phosphorylated on Ser5 and Ser2 of the C-terminal domain (CTD). In addition, Sus1 copurifies with the essential mRNA export factors Yra1 and Mex67, which bind to the mRNA cotranscriptionally. Consistently, ChIP analysis reveals that Sus1 is present at coding regions dependent on transcription in a manner stimulated by Kin28-dependent CTD phosphorylation. Strikingly, eliminating the TREX2 component Sac3 or the SAGA subunit Ubp8 partially impairs Sus1 targeting to coding sequences and upstream activating sequences (UAS). We found, unexpectedly, that Sgf73 is necessary for association of Sus1 with both SAGA and TREX2, and that its absence dramatically reduces Sus1 occupancy of UAS and ORF sequences. Our results reveal that Sus1 plays a key role in coordinating gene transcription and mRNA export by working at the interface between the SAGA and TREX2 complexes during transcription elongation.
Mesh Terms:
Blotting, Western, Chromatin Immunoprecipitation, Gene Expression Regulation, Fungal, Immunoprecipitation, Methylation, Nuclear Proteins, Open Reading Frames, Phosphoproteins, Phosphorylation, RNA Polymerase II, RNA Transport, RNA, Messenger, RNA-Binding Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Trans-Activators, Transcription, Genetic
Blotting, Western, Chromatin Immunoprecipitation, Gene Expression Regulation, Fungal, Immunoprecipitation, Methylation, Nuclear Proteins, Open Reading Frames, Phosphoproteins, Phosphorylation, RNA Polymerase II, RNA Transport, RNA, Messenger, RNA-Binding Proteins, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins, Trans-Activators, Transcription, Genetic
Genes Dev.
Date: Oct. 15, 2008
PubMed ID: 18923079
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