Summary
Transcriptional start sites of the rpoH gene which codes for a minor σ factor (σ32) of Escherichia coli RNA polymerase were determined. The rpoH gene is transcribed, both in vivo and in vitro, from two major (P1 and P2) and one minor (P2*) promoters. In vitro synthesis of the rpoH mRNAs is dependent on the major species of RNA polymerase holoenzyme (Eσ70) but not on the minor one (Eσ32). S1 nuclease analysis of the in vivo RNA showed that the level of rpoH transcript from the downstream P2 promoter increases rapidly when E. coli cells are transferred from 30° C to 42° C, while the transcript from the upstream P1 promoter remains at a constant level. Under these conditions, the metabolic stabilities of rpoH mRNAs are virtually unaffected, suggesting that the synthesis of rpoH mRNA from the P2 promoter is specifically enhanced upon heatshock.
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References
Aiba H, Adhya S, de Crombrugghe B (1981) Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem 256:11905–11910
Baker TA, Grossman AD, Gross CA (1984) A gene regulating the heat shock response in Escherichia coli also affects proteolysis. Proc Natl Acad Sci USA 81:6779–6783
Bloom M, Skelly S, VanBogelen R, Neidhardt F, Brot N, Weissbach H (1986) In vitro effect of the Escherichia coli heat shock regulatory protein on expression of heat shock genes. J Bacteriol 166:380–384
Cowing DW, Bardwell JCA, Craig EA, Woolford C, Hendrix RW, Gross CA (1985) Consensus sequence for Escherichia coli heat shock gene promoters. Proc Natl Acad Sci USA 82:2679–2683
Crickmore N, Salmond GPC (1986) The Escherichia coli heat shock regulatory gene is immediately downstream of a cell division operon: The fam mutation is allelic with rpoH. Mol Gen Genet 205:535–539
Fujita N, Nomura T, Ishihama A (1987a) Promoter selectivity of Escherichia coli RNA polymerase. Purification and properties of holoenzyme containing the heat-shock σ subunit. J Biol Chem 262:1855–1859
Fujita N, Ishihama A, Nagasawa Y, Ueda S (1987b) RNA polymerase sigma-related proteins in Escherichia coli: Detection by antibodies against a synthetic peptide. Mol Gen Genet 210:5–9
Goff SA, Goldberg AL (1985) Production of abnormal proteins in E. coli stimulates transcription of lon and other heat shock genes. Cell 41:587–595
Goff SA, Casson LP, Goldberg AL (1984) Heat shock regulatory gene htpR influences rates of protein degradation and expression of the lon gene in Escherichia coli. Proc Natl Acad Sci USA 81:6647–6651
Grossman AD, Erickson JW, Gross CA (1984) The htpR gene product of E. coli is a sigma factor for heat-shock promoters. Cell 38:383–390
Grossman AD, Zhou YN, Gross CA, Heilig J, Christie GE, Calender R (1985) Mutations in the rpoH (htpR) gene of Escherichia coli K-12 phenotypically suppress a temperature-sensitive mutant defective in the 15-1 subunit of RNA polymerase. J Bacteriol 161:939–943
Kajitani M, Ishihama A (1983a) Determination of the promoter strength in the mixed transcription system: Promoters of lactose, tryptophan and ribosomal protein L10 operons from Escherichia coli. Nucleic Acids Res 11:671–686
Kajitani M, Ishihama A (1984b) Determination of the promoter strength in the mixed transcription system, II. Promoters of ribosomal RNA, ribosomal protein S1 and recA protein operons from Escherichia coli. Nucleic Acids Res 11:3873–3888
Kajitani M, Ishihama A (1984) Promoter selectivity of Escherichia coli RNA polymerase. Differential stringent control of the multiple promoters from ribosomal RNA and protein operons. J Biol Chem 259:1951–1957
Kato A, Ishihama A, Noda A, Ueda S (1984) Improved purification and enzymatic properties of three forms of reverse transcriptase from avian myeloblastosis virus. J Virol Methods 9:325–339
Landick R, Vaughn V, Lau ET, VanBogelen RA, Erickson JW, Neidhardt FC (1984) Nucleotide sequence of the heat shock regulatory gene of E. coli suggests its protein product may be a transcription factor. Cell 38:175–182
Maxam AM, Gilbert W (1977) A new method for sequencing DNA. Proc Natl Acad Sci USA 74:560–564
Neidhardt FC, VanBogelen RA (1981) Positive regulatory gene for temperature-controlled proteins in Escherichia coli. Biochem Biophys Res Commun 100:894–900
Neidhardt FC, VanBogelen RA, Lau ET (1983) Molecular cloning and expression of a gene that controls the high-temperature regulon of Escherichia coli. J Bacteriol 153:597–603
Neidhardt FC, VanBogelen RA, Vaughn V (1984) The genetics and regulation of heat-shock proteins. Annu Rev Genet 18:295–329
Osawa T, Yura T (1981) Effects of reduced amount of RNA polymerase sigma factor on gene expression and growth of Escherichia coli: Studies of the rpoD40 (amber) mutation. Mol Gen Genet 184:166–173
Pedersen S, Skouv J, Kajitani M, Ishihama A (1984) Transcriptional organization of the rpsA operon of Escherichia coli. Mol Gen Genet 196:135–140
Piette J, Nyxtoyr H, Lusty CJ, Cunin R, Weyens G, Crabeel M, Charlier D, Glansdorff N, Pierard A (1984) DNA sequence of the carA gene and the control region of carAB: tandem promoters, respectively controlled by arginine and pyrimidines, regulate the synthesis of carbamoyl-phosphate synthetase in Escherichia coli K-12. Proc Natl Acad Sci USA 81:4134–4138
Reitzer LJ, Magasanik B (1985) Expression of glnA in Escherichia coli is regulated at tandem promoters. Proc Natl Acad Sci USA 82:1979–1983
Sancar GB, Sancar A, Little JW, Rupp WD (1982) The uvrB gene of Escherichia coli has both lexA-repressed and lexA-independent promoters. Cell 28:523–530
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Taylor WE, Straus DB, Grossman AD, Burton ZF, Gross CA, Burgess RR (1984) Transcription from a heat-inducible promoter causes heat shock regulation of the sigma subunit of E. coli RNA polymerase. Cell 38:371–381
Tilly K, Erickson J, Sharma S, Georgopoulos C (1986) Heat shock regulatory gene rpoH mRNA level increases after heat shock in Escherichia coli. J Bacteriol 168:1155–1158
Tsuchido T, VanBogelen RA, Neidhardt FC (1986) Heat shock response in Escherichia coli influences cell division. Proc Natl Acad Sci USA 83:6959–6963
Yamamori T, Yura T (1980) Temperature-induced synthesis of specific proteins in Escherichia coli: Evidence for transcriptional control. J Bacteriol 142:843–851
Yamamori T, Yura T (1982) Genetic control of heat-shock protein synthesis and its bearing on growth and thermal resistance in Escherichia coli K-12. Proc Natl Acad Sci USA 79:860–864
Yamamori T, Osawa T, Tobe T, Ito K, Yura T (1982) Escherichia coli gene (hin) controls transcription of heat-shock operons and cell growth at high temperature. In: Schlessinger MJ, Ashburner M, Tissières A (eds) Heat shock from bacteria to man. Cold Spring Harbor Laboratory Press, New York, pp 131–137
Yura T, Tobe T, Ito K, Osawa T (1984) Heat shock regulatory gene (htpR) of Escherichia coli is required for growth at high temperature but is dispensable at low temperature. Proc Natl Acad Sci USA 81:6803–6807
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Fujita, N., Ishihama, A. Heat-shock induction of RNA polymerase sigma-32 synthesis in Escherichia coli: Transcriptional control and a multiple promoter system. Mol Gen Genet 210, 10–15 (1987). https://doi.org/10.1007/BF00337752
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DOI: https://doi.org/10.1007/BF00337752