Recruitment of TREX to the transcription machinery by its direct binding to the phospho-CTD of RNA polymerase II.
Messenger RNA (mRNA) synthesis and export are tightly linked, but the molecular mechanisms of this coupling are largely unknown. In Saccharomyces cerevisiae, the conserved TREX complex couples transcription to mRNA export and mediates mRNP formation. Here, we show that TREX is recruited to the transcription machinery by direct interaction of its subcomplex THO with the serine 2-serine 5 (S2/S5) diphosphorylated CTD of RNA polymerase II. S2 and/or tyrosine 1 (Y1) phosphorylation of the CTD is required for TREX occupancy in vivo, establishing a second interaction platform necessary for TREX recruitment in addition to RNA. Genome-wide analyses show that the occupancy of THO and the TREX components Sub2 and Yra1 increases from the 5' to the 3' end of the gene in accordance with the CTD S2 phosphorylation pattern. Importantly, in a mutant strain, in which TREX is recruited to genes but does not increase towards the 3' end, the expression of long transcripts is specifically impaired. Thus, we show for the first time that a 5'-3' increase of a protein complex is essential for correct expression of the genome. In summary, we provide insight into how the phospho-code of the CTD directs mRNP formation and export through TREX recruitment.
Gene Center and Munich Center for Integrated Protein Science CIPSM at the Department of Biochemistry of the Ludwig-Maximilians-University of Munich, Munich, Germany.
Dominik M
DM
Meinel
Cornelia
C
Burkert-Kautzsch
Anja
A
Kieser
Eoghan
E
O'Duibhir
Matthias
M
Siebert
Andreas
A
Mayer
Patrick
P
Cramer
Johannes
J
Söding
Frank C P
FC
Holstege
Katja
K
Sträßer
D000251:Adenosine Triphosphatases / Q000235:genetics* / Q000378:metabolism; D046912:Multiprotein Complexes; D009687:Nuclear Proteins / Q000235:genetics*; D010766:Phosphorylation; D012319:RNA Polymerase II / Q000235:genetics*; D012333:RNA, Messenger / Q000096:biosynthesis; D016601:RNA-Binding Proteins / Q000235:genetics*; D012261:Ribonucleoproteins / Q000235:genetics* / Q000378:metabolism; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics*; D012694:Serine / Q000235:genetics; D014157:Transcription Factors / Q000235:genetics; D014158:Transcription, Genetic; D014443:Tyrosine / Q000235:genetics
D046912:Multiprotein Complexes; D009687:Nuclear Proteins; D012333:RNA, Messenger; D016601:RNA-Binding Proteins; D012261:Ribonucleoproteins; D029701:Saccharomyces cerevisiae Proteins; C405934:THO2 protein, S cerevisiae; D014157:Transcription Factors; C105887:YRA1 protein, S cerevisiae; C026452:messenger ribonucleoprotein; D014443:Tyrosine; D012694:Serine; D012319:RNA Polymerase II; D000251:Adenosine Triphosphatases; C027900:RNA-dependent ATPase