Home
  >  
Section 11
  >  
Chapter 10,024

UTP allosterically regulates transcription by Escherichia coli RNA polymerase from the bacteriophage T7 A1 promoter

Johnson, R.S.; Chester, R.E.

Journal of Molecular Biology 318(2): 305-320

2002

ISSN/ISBN: 0022-2836
PMID: 12051839
DOI: 10.1016/s0022-2836(02)00042-6
Accession: 010023710
Download citation:  
Text
  |  
BibTeX
  |  
RIS
In the case of Escherichia coli RNA polymerase, UTP at elevated concentrations suppresses terminated transcript accumulation during multiple-round transcription from a DNA construct containing the T7 A1 promoter and T(e) terminator. The step that is affected by UTP at elevated concentrations is promoter clearance. In an attempt to understand better the mechanism by which UTP regulates this step, we analyzed the effect of UTP on the formation of pppApU in the presence of only UTP and ATP. At elevated concentrations, UTP is a non-competitive inhibitor with respect to ATP in the formation of pppApU. This indicates that the effect of UTP on the formation of pppApU is mediated through an allosteric site. Moreover, the magnitude of the inhibition of pppApU formation is sufficient to account for the decrease in terminated transcript accumulation at elevated UTP concentrations. Thus, it appears that UTP modulates terminated transcript accumulation during multiple-round transcription from this DNA construct by allosteric regulation of promoter clearance at the point of transcription initiation.

PDF emailed within 0-6 h: $19.90




Related References

Johnson, R.S.; Chester, R.E. 2001: UTP allosterically regulates transcription initiation from the bacteriophage T7 A1 promoter by Escherichia coli RNA polymerase FASEB Journal 15(5): A901
Chen, J.; Gopalkrishnan, S.; Chiu, C.; Chen, A.Y.; Campbell, E.A.; Gourse, R.L.; Ross, W.; Darst, S.A. 2019: E. coli TraR allosterically regulates transcription initiation by altering RNA polymerase conformation Elife 8
Williams, K.P.; Kassavetis, G.A.; Geiduschek, E.P. 1987: Interactions of the bacteriophage T4 gene 55 product with Escherichia coli RNA polymerase: competition with Escherichia coli σ70 and release from late T4 transcription complexes following initiation Journal of Biological Chemistry 262(25): 12365-12371
Williams, K.P.; Kassavetis, G.A.; Geiduschek, E.Peter 1987: Interactions of the bacteriophage T4 gene 55 product with Escherichia coli RNA polymerase. Competition with Escherichia coli s70 and release from late T4 transcription complexes following initiation Journal of Biological Chemistry 262: 365-371
Williams, K.P.; Kassavetis, G.A.; Geiduschek, E.P. 1987: Interactions of the bacteriophage T4 gene 55 product with Escherichia coli RNA polymerase. Competition with Escherichia coli sigma 70 and release from late T4 transcription complexes following initiation Journal of Biological Chemistry 262(25): 12365-12371
Bräutigam, A.R.; Sauerbier, W. 1973: Transcription unit mapping in bacteriophage T7. I. In vivo transcription by Escherichia coli RNA polymerase Journal of Virology 12(4): 882-886
Kadesch, T.R.; Rosenberg, S.; Chamberlin, M.J. 1982: Binding of Escherichia coli RNA polymerase holoenzyme to bacteriophage T7 DNA. Measurements of binding at bacteriophage T7 promoter A1 using a template competition assay Journal of Molecular Biology 155(1): 1-29
Margolin, W.; Howe, M.M. 1990: Activation of the bacteriophage Mu lys promoter by Mu C protein requires the s70 subunit of Escherichia coli RNA polymerase Journal of Bacteriology 172: 24-9
Prosen, D.E.; Cech, C.L. 1985: Bacteriophage T7 E promoter: identification and measurement of kinetics of association with Escherichia coli RNA polymerase Biochemistry 24(9): 2219-2227
Møllegaard, N.E.; Rasmussen, P.B.; Valentin-Hansen, P.; Nielsen, P.E. 1993: Characterization of promoter recognition complexes formed by CRP and CytR for repression and by CRP and RNA polymerase for activation of transcription on the Escherichia coli deoP2 promoter Journal of Biological Chemistry 268(23): 17471-17477
Lewicki, B.T.; Margus, T.; Remme, J.; Nierhaus, K.H. 1993: Coupling of rRNA transcription and ribosomal assembly in vivo. Formation of active ribosomal subunits in Escherichia coli requires transcription of rRNA genes by host RNA polymerase which cannot be replaced by bacteriophage T7 RNA polymerase Journal of Molecular Biology 231(3): 581-593
Parks, A.R.; Court, C.; Lubkowska, L.; Jin, D.J.; Kashlev, M.; Court, D.L. 2014: Bacteriophage λ N protein inhibits transcription slippage by Escherichia coli RNA polymerase Nucleic Acids Research 42(9): 5823-5829
Hoffman, D.J.; Niyogi, S.K. 1973: RNA initiation with dinucleoside monophosphates during transcription of bacteriophage T4 DNA with RNA polymerase of Escherichia coli Proceedings of the National Academy of Sciences of the United States of America 70(2): 574-578
Johnson, R.S.; Chester, R.E. 1998: Stopped-flow kinetic analysis of the interaction of Escherichia coli RNA polymerase with the bacteriophage T7 A1 promoter Journal of Molecular Biology 283(2): 353-370
Margolin, W.; Howe, M.M. 1990: Activation of the bacteriophage Mu lys promoter by Mu C protein requires the sigma 70 subunit of Escherichia coli RNA polymerase Journal of Bacteriology 172(3): 1424-1429
Kassavetis, G.A.; Kaya, K.M.; Chamberlin, M.J. 1978: Escherichia coli RNA polymerase-rifampicin complexes bound at promoter sites block RNA chain elongation by Escherichia coli RNA polymerase and T7-specific RNA polymerase Biochemistry 17(26): 5798-5804
Strauss, H.S.; Burgess, R.R.; Record, M.T. 1980: Binding of Escherichia coli ribonucleic acid polymerase holoenzyme to a bacteriophage T7 promoter-containing fragment: evaluation of promoter binding constants as a function of solution conditions Biochemistry 19(15): 3504-3515
Stahl, S.J.; Chamberlin, M.J. 1977: An expanded transcriptional map of T7 bacteriophage. Reading of minor T7 promoter sites in vitro by Escherichia coli RNA polymerase Journal of Molecular Biology 112(4): 577-601
James, T.D.; Cashel, M.; Hinton, D.M. 2010: A mutation within the β subunit of Escherichia coli RNA polymerase impairs transcription from bacteriophage T4 middle promoters Journal of Bacteriology 192(21): 5580-5587
Rosenberg, S.; Kadesch, T.R.; Chamberlin, M.J. 1982: Binding of Escherichia coli RNA polymerase holoenzyme to bacteriophage T7 DNA. Measurements of the rate of open complex formation at T7 promoter a Journal of Molecular Biology 155(1): 31-51