Efficient release from promoter-proximal stall sites requires transcript cleavage factor TFIIS.

Uninduced heat shock genes are poised for rapid activation, with RNA polymerase II (Pol II) transcriptionally engaged, but paused or stalled, within the promoter-proximal region. Upon heat shock, this Pol II is promptly released from the promoter region and additional Pol II and transcription factors are robustly recruited to the gene. Regulation of the heat shock response relies upon factors that modify the efficiency of elongation through the initially transcribed sequence. Here, we report that Pol II is susceptible to transcription arrest within the promoter-proximal region of Drosophila hsp70 and that transcript cleavage factor TFIIS is essential for rapid induction of hsp70 RNA. Moreover, using a tandem RNAi-ChIP assay, we discovered that TFIIS is not required to establish the stalled Pol II, but that TFIIS is critical for efficient release of Pol II from the hsp70 promoter region and the subsequent recruitment of additional Pol II upon heat induction.

[1]  William J. Rice,et al.  Structure and Function of the Transcription Elongation Factor GreB Bound to Bacterial RNA Polymerase , 2003, Cell.

[2]  B. Shafer,et al.  Genetic Interactions of DST1 in Saccharomyces cerevisiae Suggest a Role of TFIIS in the Initiation-Elongation Transition , 2004, Genetics.

[3]  D. Eick,et al.  Hold back of RNA polymerase II at the transcription start site mediates down‐regulation of c‐myc in vivo. , 1992, The EMBO journal.

[4]  J. Lis,et al.  Indirect immunofluorescent labeling of Drosophila polytene chromosomes: visualizing protein interactions with chromatin in vivo. , 2004, Methods in enzymology.

[5]  H. Handa,et al.  NELF and DSIF cause promoter proximal pausing on the hsp70 promoter in Drosophila. , 2003, Genes & development.

[6]  J. Lis,et al.  Mediator, not holoenzyme, is directly recruited to the heat shock promoter by HSF upon heat shock. , 2001, Molecular cell.

[7]  O. Laptenko,et al.  Transcript cleavage factors GreA and GreB act as transient catalytic components of RNA polymerase , 2003, The EMBO journal.

[8]  A. Sluder,et al.  Fractionation of transcription factors for RNA polymerase II from Drosophila Kc cell nuclear extracts. , 1987, The Journal of biological chemistry.

[9]  Y. Ling,et al.  Subnuclear localization and Cajal body targeting of transcription elongation factor TFIIS in amphibian oocytes. , 2003, Molecular biology of the cell.

[10]  J. Roberts,et al.  Function of transcription cleavage factors GreA and GreB at a regulatory pause site. , 2000, Molecular cell.

[11]  D. Price,et al.  Mechanism of DmS-II-mediated pause suppression by Drosophila RNA polymerase II. , 1993, The Journal of biological chemistry.

[12]  J. Lis,et al.  Short transcripts of the ternary complex provide insight into RNA polymerase II elongational pausing. , 1995, Journal of molecular biology.

[13]  D. Luse,et al.  The transition from initiation to elongation by RNA polymerase II. , 1998, Cold Spring Harbor symposia on quantitative biology.

[14]  D. Luse,et al.  Promoter Clearance by RNA Polymerase II Is an Extended, Multistep Process Strongly Affected by Sequence , 2001, Molecular and Cellular Biology.

[15]  D. Price P-TEFb, a Cyclin-Dependent Kinase Controlling Elongation by RNA Polymerase II , 2000, Molecular and Cellular Biology.

[16]  D. Reines Elongation factor-dependent transcript shortening by template-engaged RNA polymerase II. , 1992, The Journal of biological chemistry.

[17]  M. Kozlov,et al.  Donation of catalytic residues to RNA polymerase active center by transcription factor Gre , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[18]  P. Labhart Transcript Cleavage in an RNA Polymerase I Elongation Complex , 1997, The Journal of Biological Chemistry.

[19]  J. Lis,et al.  RNA polymerase II pauses at the 5' end of the transcriptionally induced Drosophila hsp70 gene , 1991, Molecular and cellular biology.

[20]  M. Kashlev,et al.  Transcriptional arrest: Escherichia coli RNA polymerase translocates backward, leaving the 3' end of the RNA intact and extruded. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[21]  J. Lis,et al.  In vivo transcriptional pausing and cap formation on three Drosophila heat shock genes. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[22]  J. Lis,et al.  P-TEFb kinase recruitment and function at heat shock loci. , 2000, Genes & development.

[23]  J. Lis,et al.  Phosphorylation of RNA polymerase II C-terminal domain and transcriptional elongation , 1994, Nature.

[24]  B. Graveley,et al.  TFIIS binds to mouse RNA polymerase I and stimulates transcript elongation and hydrolytic cleavage of nascent rRNA , 1996, Molecular and General Genetics MGG.

[25]  Danny Reinberg,et al.  Recent highlights of RNA-polymerase-II-mediated transcription. , 2004, Current opinion in cell biology.

[26]  Richard A Young,et al.  Exchange of RNA polymerase II initiation and elongation factors during gene expression in vivo. , 2002, Molecular cell.

[27]  M. Chamberlin,et al.  Escherichia coli transcript cleavage factors GreA and GreB stimulate promoter escape and gene expression in vivo and in vitro. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[28]  D. Gilmour,et al.  Analyses of promoter-proximal pausing by RNA polymerase II on the hsp70 heat shock gene promoter in a Drosophila nuclear extract , 1996, Molecular and cellular biology.

[29]  D. Luse,et al.  The RNA polymerase II ternary complex cleaves the nascent transcript in a 3'----5' direction in the presence of elongation factor SII. , 1992, Genes & development.

[30]  D. Price,et al.  Control of formation of two distinct classes of RNA polymerase II elongation complexes , 1992, Molecular and cellular biology.

[31]  D. Eick,et al.  Elongation and premature termination of transcripts initiated from c-fos and c-myc promoters show dissimilar patterns. , 1995, Oncogene.

[32]  D. Reines,et al.  Transcription elongation factor SII , 2000, BioEssays : news and reviews in molecular, cellular and developmental biology.

[33]  A. Rougvie,et al.  The RNA polymerase II molecule at the 5′ end of the uninduced hsp70 gene of D. melanogaster is transcriptionally engaged , 1988, Cell.

[34]  S. Borukhov,et al.  Transcript cleavage factors from E. coli , 1993, Cell.

[35]  J. Lis,et al.  Coordination of transcription, RNA processing, and surveillance by P-TEFb kinase on heat shock genes. , 2004, Molecular cell.

[36]  J. Lis,et al.  Transcription Factor and Polymerase Recruitment, Modification, and Movement on dhsp70 In Vivo in the Minutes following Heat Shock , 2003, Molecular and Cellular Biology.

[37]  J. Lis,et al.  The RNA processing exosome is linked to elongating RNA polymerase II in Drosophila , 2002, Nature.

[38]  C. Kane,et al.  Running with RNA polymerase: eukaryotic transcript elongation. , 2003, Trends in genetics : TIG.

[39]  C. Kane,et al.  Promoting elongation with transcript cleavage stimulatory factors. , 2002, Biochimica et biophysica acta.

[40]  A. Shilatifard,et al.  The RNA polymerase II elongation complex. , 2003, Annual review of biochemistry.

[41]  M. Mathews,et al.  HIV-1 Tat overcomes inefficient transcriptional elongation in vitro. , 1993, Journal of molecular biology.

[42]  M. Groudine,et al.  The block to transcriptional elongation within the human c-myc gene is determined in the promoter-proximal region. , 1992, Genes & development.

[43]  Y. Schwartz,et al.  Polycomb silencing blocks transcription initiation. , 2004, Molecular cell.

[44]  J. Lis,et al.  High-resolution localization of Drosophila Spt5 and Spt6 at heat shock genes in vivo: roles in promoter proximal pausing and transcription elongation. , 2000, Genes & development.

[45]  J. Lis,et al.  Promoter-associated pausing in promoter architecture and postinitiation transcriptional regulation. , 1998, Cold Spring Harbor symposia on quantitative biology.

[46]  J. Lis,et al.  Cdk7 Is Required for Full Activation of Drosophila Heat Shock Genes and RNA Polymerase II Phosphorylation In Vivo , 2003, Molecular and Cellular Biology.

[47]  Jeffrey W. Roberts,et al.  Antitermination by bacteriophage lambda Q protein. , 1998, Cold Spring Harbor symposia on quantitative biology.

[48]  P. Cramer,et al.  Architecture of the RNA Polymerase II-TFIIS Complex and Implications for mRNA Cleavage , 2003, Cell.

[49]  G. Hartzog,et al.  Transcription elongation by RNA polymerase II. , 2003, Current opinion in genetics & development.

[50]  D. Reinberg,et al.  Tracking FACT and the RNA Polymerase II Elongation Complex Through Chromatin in Vivo , 2003, Science.