Skip to main content

RNA Chain Initiation and Promoter Escape by RNA Polymerase

  • Chapter
Regulation of Gene Expression in Escherichia coli

Abstract

The efficiency of promoter function is determined by two distinct phases of transcription, promoter binding and activation, and RNA chain initiation and promoter escape. The former process has been extensively studied in both prokaryotic and eukaryotic organisms, and involves interaction of the RNA polymerase with general transcription factors, promoter specific factors, and with the DNA sequences in the recognition region of the promoter (for reviews see refs. 1–4). The latter process has generally been overlooked, and is often subtended into the promoter binding phase by use of the general term “initiation” to include both promoter binding and RNA chain initiation. However, it has been recognized for over 25 years that the two phases are distinct. Hence, true RNA chain initiation has, in some ways, become the lost step in transcription. The promoter binding and activation process can normally be completed in the absence of the nucleoside triphosphate substrates.5 Similarly, the RNA chain initiation and promoter escape phase is a biochemically distinct process, and is controlled and regulated by DNA sequences and factors that have no similar role in promoter binding. These sequences and factors can affect promoter strength considerably,6,7 and can lead to regulation that is independent of the promoter binding step.8–11

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. McClure WR. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem 1985; 54:171–204.

    Article  Google Scholar 

  2. Hoopes BC, McClure WR. Strategy in Regulation of Transcription Initiation. In: Neidhardt FC et al, eds. Escherichia coli and Salmonella typhimurium. ASM Press 1987:1231–1240.

    Google Scholar 

  3. Tjian R, Maniatis T. Transcriptional activation: a complex puzzle with few easy pieces. Cell 1994; 77:5–8.

    Article  Google Scholar 

  4. Gralla JD. Promoter recognition and mRNA initiation by Escherichia coli σ70. Methods in Enzymol 1990; 185:37–54.

    Article  Google Scholar 

  5. Chamberlin MJ. Transcription 1970: a summary. CSHSQB 1970; 35:851–873.

    Google Scholar 

  6. Kammerer W, Deuschle U, Gentz R, Bujard H. Functional dissection of Escherichia coli promoters: information in the transcribed region is involved in late steps of the overall process. EMBO J 1986; 11:2995–3000.

    Google Scholar 

  7. Knaus R, Bujard H. PL of coliphage lambda: an alternative solution for an efficient promoter. EMBO J 1988; 7:2919–2923.

    Google Scholar 

  8. Menendez M, Kolb A, Buc H. A new target for CRP action at the malT promoter. EMBO J 1987; 6:4227–4234.

    Google Scholar 

  9. Jin DJ, Turnbough CL, Jr. An Escherichia coli RNA polymerase defective in transcription due to its overproduction of abortive initiation products. J Mol Biol 1994; 236:72–80.

    Article  Google Scholar 

  10. Jin DJ. Slippage synthesis at the gal P2 promoter of Escherichia coli and its regulation by UTP concentration and cAMP.cAMP receptor protein. J Biol Chem 1994; 269:17221–17227.

    Google Scholar 

  11. Maxon ME, Goodrich JA, Tjian R. Transcription factor HE binds preferentially to RNA polymerase IIa and recruits TFIIH: a model for promoter clearance. Genes and Dev 1994; 8:515–524.

    Article  Google Scholar 

  12. McClure WR, Cech CL. On the mechanism of rifampicin inhibition of RNA synthesis. J Biol Chem 1978; 253:8949–8956.

    Google Scholar 

  13. Carpousis AJ, Gralla JD. Cycling of ribonucleic acid polymerase to produce oligonucleotides during initiation in vitro at the lac UV5 promoter. Biochemistry 1980; 19:3245–3253.

    Article  Google Scholar 

  14. Munson LM, Reznikoff WS. Abortive initiation and long ribonucleic acid synthesis. Biochemistry 1981; 20:2081–2085.

    Article  Google Scholar 

  15. Hsu LM, Chamberlin MJ. Abortive initiation, productive initiation and promoter escape by E. coli RNA polymerase. (In preparation).

    Google Scholar 

  16. Krummel B, Chamberlin MJ. RNA chain initiation by Escherichia coli RNA polymerase. Structural transitions of the enzyme in early ternary complexes. Biochemistry 1989; 28:7829–7842.

    Article  Google Scholar 

  17. Martin CT, Muller DK, Coleman JE. Processivity in early stages of transcription by T7 RNA polymerase. Biochemistry 1988; 27:3966–3974.

    Article  Google Scholar 

  18. Whipple FW, Sonenshein AL. Mechanism of initiation of transcription by Bacillus subtilis RNA polymerase at several promoters. J Mol Biol 1992; 223:399–414.

    Article  Google Scholar 

  19. Kainz M, Roberts JW. Initiation and pausing at the lambda late gene promoter in vivo. J Mol Biol 1995; 254:808–814.

    Article  Google Scholar 

  20. Ellinger T, Behnke D, Bujard H, Gralla JD. Stalling of E. coli RNA polymerase in the +6 to +12 region in vivo is associated with tight binding to consensus promoter elements. J Mol Biol 1994; 239:455–465.

    Article  Google Scholar 

  21. Grachev MA, Zaychikov EF. Initiation by Escherichia coli RNA polymerase: transformation of abortive to productive complex. FEBS Lett 1980; 115:23–26.

    Article  Google Scholar 

  22. Carpousis AJ, Gralla JD. Interaction of RNA polymerase with lac UV5 promoter DNA during mRNA initiation and elongation. Footprinting, methylation and rifampicin-sensitivity changes accompanying transcription initiation. J Mol Biol 1985; 183:165–177.

    Article  Google Scholar 

  23. Straney DC, Crothers DM. A stressed intermediate in the formation of stably initiated RNA chains at the Escherichia coli lac UV5 promoter. J Mol Biol 1987; 193:279–292.

    Article  Google Scholar 

  24. Hansen UM, McClure WR. Role of a subunit of Escherichia coli RNA polymerase in initiation. II. Release of a from ternary complexes. J Biol Chem 1980; 255:9564–9570.

    Google Scholar 

  25. Schultz W, Zillig W. Nucleic Acids Res 1981; 9:6889–6906.

    Article  Google Scholar 

  26. Metzger W, Schickor P, Meier T et al. Nucleation of RNA chain formation by Escherichia coli DNA-dependent RNA polymerase. J Mol Biol 1993; 232:35–49.

    Article  Google Scholar 

  27. Borukhov S, Sagitov V, Josaitis CA et al. Two modes of transcription initiation in vitro at the rrnB P1 promoter of Escherichia coli. J Biol Chem 1993; 268:23477–23482.

    Google Scholar 

  28. Severinov K, Goldfarb A. Topology of the product binding site in RNA polymerase revealed by transcript slippage at the phage λ PL promoter. J Biol Chem 1994; 269:31701–31705.

    Google Scholar 

  29. Ruetsch NR, Dennis D. RNA polymerase. Limit cognate primer for initiation and stable ternary complex fromation. J Biol Chem 1987; 262:1674–1679.

    Google Scholar 

  30. Milligan JF, Groebe DR, Witherell GW, Uhlenbeck OC. Oligonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res 1987; 15:8783–8798.

    Article  Google Scholar 

  31. Nam S-C, Kang C. Transcription initiation site selection and abortive initiation: cycling of phage SP6 RNA polymerase. J Biol Chem 1988; 263:18123–18127.

    Google Scholar 

  32. Ling M-L, Risman SS, Klement JF et al. Abortive initiation by bacteriophage T3 and T7 RNA polymerases under conditions of limiting substrate. Nucleic Acids Res 1989; 17:1605–1618.

    Article  Google Scholar 

  33. Hinrichsen AI, Ortner I, Hartmann GR. Synthesis of dinucleoside tetraphosphates by RNA polymerase B (II) from calf thymus. FEBS Lett 1985; 193:199–202.

    Article  Google Scholar 

  34. Mosig H, Schaffner AR, Sieber H, Hartmann GR. Primer-independent abortive initiation by wheat germ RNA polymerase B (II). Eur J Biochem 1985; 149:337–343.

    Article  Google Scholar 

  35. Luse DS, Jacob GA. Abortive initiation by RNA polymerase II in vitro at the adenovirus 2 major late promoter. J Biol Chem 1987; 262: 14990–14997.

    Google Scholar 

  36. Goodrich JA, Tjian R. Transcription factor IIE, IIH and ATP hydrolysis direct promoter clearance by RNA polymerase II. Cell 1994; 77:145–156.

    Article  Google Scholar 

  37. Johnston DE, McClure WR. Abortive initiation of in vitro RNA synthesis on bacteriophage λ DNA. In: Losick RR, Chamberlin MJ, eds. RNA Polymerase. Cold Spring Harbor Laboratory, 1976:101–126.

    Google Scholar 

  38. Gralla JD, Carpousis AJ, Stefano JE. Productive and abortive initiation of transcription in vitro at the lac UV5 promoter. Biochemistry 1980; 19:5864–5869.

    Article  Google Scholar 

  39. Harley CB, Lawrie J, Boyer HW, Hedgpeth J. Reiterative copying by E. coli RNA polymerase during transcription initiation of mutant pBR322 tet promoters. Nucleic Acids Res 1990; 18:547–552.

    Article  Google Scholar 

  40. Jacques J-P, Susskind MM. Pseudo-templated transcription by Escherichia coli RNA polymerase at a mutant promoter. Genes Dev 1990; 4: 1801–1810.

    Article  Google Scholar 

  41. Guo H-C, Roberts JW. Heterogeneous initiation due to slippage at the bacteriophage 82 late gene promoter in vitro. Biochemistry 1990; 29:10702–10709.

    Article  Google Scholar 

  42. Xiong XF, Reznikoff WS. Transcriptional slippage during the transcription initiation process at a mutant lac promoter in vivo. J Mol Biol 1993; 231:569–580.

    Article  Google Scholar 

  43. Chamberlin M, Berg P. Deoxyribonucleic acid-directed synthesis of ribonucleic acid by an enzyme from Escherichia coli. Proc Natl Acad Sci USA 1962; 48:81–94.

    Article  Google Scholar 

  44. Chamberlin M, Berg P. Mechanism of RNA polymerase action: characterization of the DNA-dependent synthesis of polyadenylic acid. J Mol Biol 1964; 8:708–726.

    Article  Google Scholar 

  45. Falaschi A, Adler J, Khorana HG. Chemically synthesized deoxypoly-nucleotides as templates for ribonucleic acid polymerase. J Biol Chem 1963; 238:3080–3085.

    Google Scholar 

  46. . Hawley DK, McClure WR. Compilation and analysis of Escherichia coli promoter DNA sequences. Nucleic Acids Res 1983; 8:2237–2255.

    Article  Google Scholar 

  47. Lisser S, Margalit H. Compilation of E. coli mRNA promoter sequences. Nucleic Acids Res 1993; 21:1507–1516.

    Article  Google Scholar 

  48. Feng GH, Lee DN, Wang D et al. Gre-A induced transcript cleavage in transcription complexes containing Escherichia coli RNA polymerase is controlled by multiple factors, including nascent transcript location and structure. J Biol Chem 1994; 269:22282–22294.

    Google Scholar 

  49. MacDonald LE, Zhou Y, McAllister WT. Termination and slippage by bacteriophage T7 RNA polymerase. J Mol Biol 1993; 232:1030–1047.

    Article  Google Scholar 

  50. Wagner LA, Weiss RB, Driscoll R et al. Transcription slippage occurs during elongation at runs of adenine or thymine in Escherichia coli. Nucleic Acids Res 1990; 18:3529–3535.

    Article  Google Scholar 

  51. Zhou, Reines, Doetsch. T7 RNA polymerase bypass of large gaps on the template strand reveals a critical role of the nontemplate strand in elongation. Cell 1995; 82:577–585.

    Article  Google Scholar 

  52. Levin JR, Chamberlin MJ. Mapping and characterization of transcriptional pause sites in the early genetic region of bacteriophage T7. J Mol Biol 1987; 196:61–84.

    Article  Google Scholar 

  53. Bernhard SL, Meares CF. The a subunit of RNA polymerase contacts the leading ends of transcripts 9–13 bases long on the λ PR promoter but not on T7 A1. Biochemistry 1986; 25:5914–5919.

    Article  Google Scholar 

  54. Nierman WC, Chamberlin MJ. Studies of RNA chain initiation by Escherichia coli RNA polymerase bound to T7 DNA. Direct analysis of the kinetics of RNA chain initiation at T7 promoter A1. J Biol Chem 1979; 254:7921–7926.

    Google Scholar 

  55. Nierman WC, Chamberlin MJ. Studies of RNA chain initiation by Escherichia coli RNA polymerase bound to T7 DNA. Direct analysis of the kinetics of RNA chain initiation at T7 promoter A2. J Biol Chem 1980; 255:1819–1823.

    Google Scholar 

  56. Stefano JE, Gralla J. Lac UV5 transcription in vitro. Rate limitation subsequent to formation of an RNA polymerase-DNA complex. Biochemistry 1979; 18:1063–1067.

    Article  Google Scholar 

  57. Surratt CK, Milan SC, Chamberlin MJ. Spontaneous cleavage of RNA in ternary complexes of Escherichia coli RNA polymerase and its significance for the mechanism of transcription. Proc Natl Acad Sci USA 1991; 88:7983–7987.

    Article  Google Scholar 

  58. Levin JR, Krummel B, Chamberlin MJ. Isolation and properties of transcribing ternary complexes of Escherichia coli RNA polymerase positioned at a single template bases. J Mol Biol 1987; 196:85–100.

    Article  Google Scholar 

  59. Arndt KM, Chamberlin MJ. Transcription termination in Escherichia coli. Measurement of the rate of enzyme release from Rho-independent terminators. J Mol Biol 1988; 202:271–285.

    Article  Google Scholar 

  60. Deuschle U, Kammerer W, Gentz R, Bujard H. EMBO J 1986; 5:2987–2994.

    Google Scholar 

  61. Brunner M, Bujard H. Promoter recognition and promoter strength in the Escherichia coli system. EMBO J 1987; 6:3139–3144.

    Google Scholar 

  62. Hsu LM, Vo NV, Chamberlin MJ. Escherichia coli transcript cleavage factors GreA and GreB stimulate promoter clearance and gene expression in vitro and in vivo. Proc Natl Acad Sci USA 1995; 92:11588–11592.

    Article  Google Scholar 

  63. Kashlev M, Lee J, Zalenskaya K et al. Blocking of the initiation-to-elongation transition by a transdominant RNA polymerase mutation. Science 1990; 248:1006–1009.

    Article  Google Scholar 

  64. . Lee J, Kashlev M, Borukhov S, Goldfarb A. A beta subunit mutation disrupting the catalytic function of Escherichia coli RNA polymerase. Proc Natl Acad Sci USA 1991; 88:6018–6022.

    Article  Google Scholar 

  65. Sagitov V, Nikiforov V, Goldfarb A. Dominant lethal mutations near the 5’ substrate binding site affect RNA polymerase propagation. J Biol Chem 1993; 268:2195–2202.

    Google Scholar 

  66. . Straney S, Crothers DM. Lac repressor is a transient gene-activating protein. Cell 1987; 51:699–707.

    Article  Google Scholar 

  67. . Lee J, Goldfarb A. Lac repressor acts by modifying the initial transcribing complex so that it cannot leave the promoter. Cell 1991; 66:793–798.

    Article  Google Scholar 

  68. Muller DK, Martin CT, Coleman JE. Processivity of proteolytically modified forms of T7 RNA polymerase. Biochemistry 1988; 27:5763–5771.

    Article  Google Scholar 

  69. Chamberlin M. The selectivity of transcription. Annu Rev Biochem 1974; 43:721–775.

    Article  Google Scholar 

  70. Reynolds R, Bermudez-Cruz RM, Chamberlin MJ. Parameters affecting transcription termination by Escherichia coli RNA polymerase. I. Analysis of 13 rho-independent terminators. J Mol Biol 1992; 224:31–51.

    Article  Google Scholar 

  71. Leirmo S, Harrison C, Cayley DS, Burgess RR, Records MT, Jr. Replacement of potassium chloride by potassium glutamate dramatically enhances protein-DNA interactions in vitro. Biochemistry 1987; 26: 2095–2101.

    Article  Google Scholar 

  72. Gamper HB, Hearst JE. A topological model for transcription based on unwinding angle analysis of E. coli RNA polymerase binary, initiation and ternary complexes. Cell 1982; 29:81–90.

    Article  Google Scholar 

  73. von Hippel PH, Bear DG, Morgan WD, McSwiggen JA. Protein nucleic acid interaction in transcription: a molecular analysis. Annu Rev Biochem 1984; 53:389–446.

    Article  Google Scholar 

  74. . Yager TD, von Hippel PH. Transcription elongation and termination in Escherichia coli. In: Neidhardt FC et al, eds. Escherichia coli and Salmonella typhimurium. ASM Press 1987:1241–1275.

    Google Scholar 

  75. Rice GA, Kane CM, Chamberlin MJ. Footprinting analysis of mammalian RNA polymerase II along its transcript: an alternative view of transcript elongation. Proc Natl Acad Sci USA 1991; 88:4245–4249.

    Article  Google Scholar 

  76. . Chamberlin MJ. New models for the mechanism of transcription elongation and its regulation. Harvey Lectures, Series 88, New York: Wiley-Lis, 1995:1–21.

    Google Scholar 

  77. . Milan S, Chamberlin MJ. Structural analysis of ternary complexes of Escherichia coli RNA polymerase. Ribonuclease footprinting of nascent transcripts. 1995; (in preparation).

    Google Scholar 

  78. von Hippel PH, Yager TD. Transcript elongation and termination are competitive kinetic processes. Proc Natl Acad Sci USA 1991; 88: 2307–2311.

    Article  Google Scholar 

  79. Johnson TL, Chamberlin MJ. Complexes of yeast RNA Polymerase II and RNA are substrates for TFIIS-induced RNA cleavage. Cell 1994; 77:217–224.

    Article  Google Scholar 

  80. Nudler E, Goldfarb A, Kashlev M. Discontinuous mechanism of transcription elongation. Science 1994; 265:793–796.

    Article  Google Scholar 

  81. Chan CL, Landick R. New perspectives on RNA chain elongation and termination by E. coli RNA polymerase. In: Conaway RC, Conaway JW, eds. Transcription: Mechanisms and Regulation. New York: Raven Press, Ltd., 1994:297–321.

    Google Scholar 

  82. Wang D, Meier T, Chan C et al. Discontinuous movements of DNA and RNA in RNA polymerase accompany formation of a paused transcription complex. Cell 1995; 81:341–350.

    Article  Google Scholar 

  83. Altmann CR, Solow-Cordero DE, Chamberlin MJ. RNA cleavage and chain elongation by Escherichia coli DNA-dependent RNA polymerase in a binary enzyme-RNA complex. Proc Natl Acad Sci USA 1994; 91:3784–3788.

    Article  Google Scholar 

  84. Rudd MD, Izban MG, Luse DL. The active site of RNA polymerase II participates in transcript cleavage within arrested ternary complexes. Proc Natl Acad Sci USA 1994; 91:8057–8061.

    Article  Google Scholar 

  85. Borukhov S, Polyakov A, Nikiforov V, Goldfarb A. GreA protein: a transcription elongation factor from Escherichia coli. Proc Natl Acad Sci USA 1992; 89:8899–8902.

    Article  Google Scholar 

  86. Borukhov S, Sagitov V, Goldfarb A. Transcript cleavage factor from E. coli. Cell 1993; 72:459–466.

    Article  Google Scholar 

  87. Krummel B, Chamberlin MJ. Structural analysis of ternary complexes of Escherichia coli RNA polymerase. Individual complexes halted along different transcription units have distinct and unexpected biochemical properties. J Mol Biol 1992; 225:221–237.

    Article  Google Scholar 

  88. Krummel B, Chamberlin MJ. Structural analysis of ternary complexes of Escherichia coli RNA polymerase. Deoxyribonuclease I footprint of defined complexes. J Mol Biol 1992; 225:239–250.

    Article  Google Scholar 

  89. Mustaev A, Kashlev M, Zaychikov et al. Active center rearrangement in RNA polymerase initiation complex. J Biol Chem 1993; 268:19185–19187.

    Google Scholar 

  90. Mustaev A, Zaychikov E, Severinov K et al. Topology of the RNA polymerase active center probed by chimeric rifampicin-nucleotide compounds. Proc Natl Acad Sci USA 1994; 91:12036–12040.

    Article  Google Scholar 

  91. Ross W, Gosink KK, Salomon J et al. A third recognition element in bacterial promoters: DNA binding by the a subunit of RNA polymerase. Science 1993; 262:1407–1413.

    Article  Google Scholar 

  92. Krakow JS, von der Helm K. Azotobacter RNA polymerase transitions and the release of sigma. Cold Spring Harbor Symp. Quant Biol 1970; 35:73–82.

    Article  Google Scholar 

  93. Busby S, Spaasky A, Buc H. On the binding of tRNA to Escherichia coli RNA polymerase. Interactions between the core enzyme, DNA and tRNA. Eur J Biochem 1981; 118:443–451.

    Article  Google Scholar 

  94. Grachev M, Kolocheva T, Lukhtanov E et al. Studies on the functional topography of Escherichia coli RNA polymerase. Highly selective affinity labeling by analogs of initiating substrates. Eur J Biochem 1987; 163:113–121

    Article  Google Scholar 

  95. Orlova M, Newlands J, Das A et al. Intrinsic cleavage activity of RNA polymerase. Proc Natl Acad Sci USA 1995; 92: 4596–4600.

    Article  Google Scholar 

Download references

Authors

Rights and permissions

Reprints and permissions

Copyright information

© 1996 R.G. Landes Company

About this chapter

Cite this chapter

Chamberlin, M.J., Hsu, L.M. (1996). RNA Chain Initiation and Promoter Escape by RNA Polymerase. In: Regulation of Gene Expression in Escherichia coli . Springer, Boston, MA. https://doi.org/10.1007/978-1-4684-8601-8_2

Download citation

  • DOI: https://doi.org/10.1007/978-1-4684-8601-8_2

  • Publisher Name: Springer, Boston, MA

  • Print ISBN: 978-1-4684-8603-2

  • Online ISBN: 978-1-4684-8601-8

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics