Crosstalk between mRNA 3' end processing and transcription initiation.

Transcription and mRNA maturation are interdependent events. Although stimulatory connections between these processes within the same round of transcription are well described, functional coupling between separate transcription cycles remains elusive. Comparing time-resolved transcription profiles of single-copy integrated β-globin gene variants, we demonstrate that a polyadenylation site mutation decreases transcription initiation of the same gene. Upon depletion of the 3' end processing and transcription termination factor PCF11, endogenous genes exhibit a similar phenotype. Readthrough RNA polymerase II (RNAPII) engaged on polyadenylation site-mutated transcription units sequester the transcription initiation/elongation factors TBP, TFIIB and CDK9, leading to their depletion at the promoter. Additionally, high levels of TBP and TFIIB appear inside the gene body, and Ser2-phosphorylated RNAPII accumulates at the promoter. Our data demonstrate that 3' end formation stimulates transcription initiation and suggest that coordinated recycling of factors from a gene terminator back to the promoter is essential for sustaining continued transcription.

[1]  Mikkel H. Schierup,et al.  RNA Exosome Depletion Reveals Transcription Upstream of Active Human Promoters , 2008, Science.

[2]  E. Cho,et al.  Phosphorylation of the Yeast Rpb1 C-terminal Domain at Serines 2, 5, and 7* , 2009, The Journal of Biological Chemistry.

[3]  S. Buratowski.  Progression through the RNA polymerase II CTD cycle. , 2009, Molecular cell.

[4]  N. Proudfoot,et al.  Pause Sites Promote Transcriptional Termination of Mammalian RNA Polymerase II , 2006, Molecular and Cellular Biology.

[5]  D. Bentley,et al.  Capping, splicing, and 3' processing are independently stimulated by RNA polymerase II: different functions for different segments of the CTD. , 2001, Genes & development.

[6]  N. Proudfoot.  How RNA polymerase II terminates transcription in higher eukaryotes. , 1989, Trends in biochemical sciences.

[7]  N. Proudfoot,et al.  Molecular Dissection of Mammalian RNA Polymerase II Transcriptional Termination , 2008, Molecular cell.

[8]  R. Palmiter,et al.  Introns increase transcriptional efficiency in transgenic mice. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[9]  M. Hampsey,et al.  A physiological role for gene loops in yeast. , 2009, Genes & development.

[10]  L. Tora,et al.  Human U1 snRNA forms a new chromatin-associated snRNP with TAF15 , 2009, EMBO reports.

[11]  D. Price,et al.  Flavopiridol Inactivates P-TEFb and Blocks Most RNA Polymerase II Transcription in Vivo * , 2001, The Journal of Biological Chemistry.

[12]  Hiroshi Kimura,et al.  U1 snRNA associates with TFIIH and regulates transcriptional initiation , 2002, Nature Structural Biology.

[13]  E. Cho,et al.  Different phosphorylated forms of RNA polymerase II and associated mRNA processing factors during transcription. , 2000, Genes & development.

[14]  N. Proudfoot,et al.  Human Pcf11 enhances degradation of RNA polymerase II-associated nascent RNA and transcriptional termination , 2007, Nucleic acids research.

[15]  Xiang-Dong Fu,et al.  Functional integration of transcriptional and RNA processing machineries. , 2008, Current opinion in cell biology.

[16]  A. Furger,et al.  Promoter proximal splice sites enhance transcription. , 2002, Genes & development.

[17]  M. Keogh,et al.  Ctk1 promotes dissociation of basal transcription factors from elongating RNA polymerase II , 2009, The EMBO journal.

[18]  T. Maniatis,et al.  An extensive network of coupling among gene expression machines , 2002, Nature.

[19]  Michael Hampsey,et al.  A role for the CPF 3'-end processing machinery in RNAP II-dependent gene looping. , 2005, Genes & development.

[20]  Frédéric Devaux,et al.  THO/Sub2p Functions to Coordinate 3′-End Processing with Gene-Nuclear Pore Association , 2008, Cell.

[21]  Xiang-Dong Fu,et al.  The splicing factor SC35 has an active role in transcriptional elongation , 2008, Nature Structural &Molecular Biology.

[22]  D. Bentley,et al.  "Cotranscriptionality": the transcription elongation complex as a nexus for nuclear transactions. , 2009, Molecular cell.

[23]  N. Proudfoot,et al.  Human 5′ → 3′ exonuclease Xrn2 promotes transcription termination at co-transcriptional cleavage sites , 2004, Nature.

[24]  X. Darzacq,et al.  In vivo dynamics of RNA polymerase II transcription , 2007, Nature Structural &Molecular Biology.

[25]  Antonin Morillon,et al.  Gene loops juxtapose promoters and terminators in yeast , 2004, Nature Genetics.

[26]  Hiroshi Handa,et al.  P-TEFb-mediated phosphorylation of hSpt5 C-terminal repeats is critical for processive transcription elongation. , 2006, Molecular cell.

[27]  O. Rozenblatt-Rosen,et al.  The multifunctional protein p54nrb/PSF recruits the exonuclease XRN2 to facilitate pre-mRNA 3' processing and transcription termination. , 2007, Genes & development.

[28]  S. Buratowski,et al.  Phosphorylation of serine 2 within the RNA polymerase II C-terminal domain couples transcription and 3' end processing. , 2004, Molecular cell.

[29]  Jesse C. Gatlin,et al.  Ribozyme cleavage reveals connections between mRNA release from the site of transcription and pre-mRNA processing. , 2005, Molecular cell.

[30]  N. Proudfoot,et al.  Gene loops function to maintain transcriptional memory through interaction with the nuclear pore complex. , 2009, Genes & development.

[31]  Melissa J. Moore,et al.  Pre-mRNA Processing Reaches Back toTranscription and Ahead to Translation , 2009, Cell.

[32]  T. Jensen,et al.  SMG6 promotes endonucleolytic cleavage of nonsense mRNA in human cells , 2009, Nature Structural &Molecular Biology.

[33]  Patricia Richard,et al.  Transcription termination by nuclear RNA polymerases. , 2009, Genes & development.

[34]  D. Bentley,et al.  RNA polymerase II pauses and associates with pre-mRNA processing factors at both ends of genes , 2008, Nature Structural &Molecular Biology.

[35]  N. Krogan,et al.  The yeast Rat1 exonuclease promotes transcription termination by RNA polymerase II , 2004, Nature.

[36]  J. Dantonel,et al.  Transcription factor TFIID recruits factor CPSF for formation of 3′ end of mRNA , 1997, Nature.

[37]  Stavroula Mili,et al.  Distinct RNP Complexes of Shuttling hnRNP Proteins with Pre-mRNA and mRNA: Candidate Intermediates in Formation and Export of mRNA , 2001, Molecular and Cellular Biology.

[38]  Arlen W. Johnson,et al.  The role of Rat1 in coupling mRNA 3'-end processing to transcription termination: implications for a unified allosteric-torpedo model. , 2006, Genes & development.

[39]  Watt W Webb,et al.  Intranuclear distribution and local dynamics of RNA polymerase II during transcription activation. , 2007, Molecular cell.

[40]  M. Hampsey,et al.  A transcription-independent role for TFIIB in gene looping. , 2007, Molecular cell.

[41]  N. Proudfoot,et al.  Transcriptional Termination Enhances Protein Expression in Human Cells , 2009, Molecular cell.

[42]  D. Bentley,et al.  Dynamic association of capping enzymes with transcribing RNA polymerase II. , 2000, Genes & development.

[43]  J. Kjems,et al.  A 5' splice site enhances the recruitment of basal transcription initiation factors in vivo. , 2008, Molecules and Cells.

[44]  S. Berget,et al.  Mutation of the AAUAAA polyadenylation signal depresses in vitro splicing of proximal but not distal introns. , 1991, Genes & development.