Summary
The Bacillus subtilis veg promotor site was shown to be composed of two neighboring RNA polymerase binding sites, only one of which (“Site I”) produces a transcript. The relationship between these sites has been investigated following (a) insertion of ten base pairs of DNA between the sites and subsequent cloning of each site independent of the other, and (b) deletion of DNA sequences upstream from the productive site. The effect of DNA insertion was to permit transcription initiation from the previously nonproductive Site II, in either the presence or absence of Site I. This could be attributed to insertion of a purine residue seven base pairs dowstream from the Pribnow box of Site II. Transcription from Site I was slightly increased in the presence of Site II cis, but was still efficient in the absence of Site II, indicating that there is no absolute dependence on Site II for in vitro transcription from Site I.
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References
Banner CD, Moran CP Jr, Losick R (1983) Deletion analysis of a complex promoter for a developmentally regulated gene from Bacillus subtilis. J Mol Biol 168:351–365
Backman K, Ptashne M, Gilbert W (1976) Construction of plasmids carrying the cI gene of bacteriophage λ. Proc Natl Acad Sci USA 73:4174–4178
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW, Crosa JG, Falkow S (1977) Construction and characterization of new cloning vehicles, II. A multipurpose cloning system. Gene 2:95–113
Gilman MZ, Chamberlin MJ (1983) Developmental and genetic regulation of Bacillus subtilis genes transcribed by sigma-28 RNA polymerase. Cell 35:285–293
Hawley D, McClure W (1983) Compilation and analysis of E. coli promoter sequences. Nucl Acids Res 11:2237–2255
Kajitani M, Ishihama A (1983) 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. Nucl Acids Res 11:3873–3888
Le Grice SFJ, Sonenshein AL (1982) Interaction of Bacillus subtilis RNA polymerase with a chromosomal promoter. J Mol Biol 162:551–564
Mackie GA, Pearsons GD (1983) Tandem promoters in the gene for ribosomal protein S20. J Biol Chem 258:7840–7846
Malan TP, McClure W (1984) Dual promoter control of the Escherichia coli lactose operon. Cell 39:173–180
Mandecki W, Reznikoff W (1982) A lac promoter with a changed distance between-10 and-35 regions. Nucl Acids Res 10:903–912
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. In: Grossman L, Moldave K (eds) Methods in enzymology, vol 65. Academic Press, New York, pp 499–560
Moran CP Jr, Lang N, Le Grice SFJ, Stephens M, Lee G, Pero J, Sonenshein AL, Losick R (1982) Nucleotide sequences that signal initiation of transcription and translation in Bacillus subtilis. Mol Gen Genet 186:339–346
Mulligan M, Brosius J, McClure W (1985) Characterization in vitro of the effect of spacer length on the activity of E. coli RNA polymerase at the tac promoter. J Biol Chem 260:3529–3538
Murray CL, Rabinowitz JC (1982) Nucleotide sequences of transcription and translation initiation regions in Bacillus phage ϕ 29 early genes. J Biol Chem 257:1053–1062
Ollington JF, Haldenwang WG, Huynh TV, Losick R (1981) Developmentally regulated transcription in a cloned segment of the Bacillus subtilis chromosome. J Bacteriol 147:432–442
Peterson ML, Reznikoff WS (1985) Properties of lac P2 in vivo and in vitro. An overlapping RNA polymerase binding site within the lactose promoter. J Mol Biol 185:535–543
Pribnow D (1975) Nucleotide sequence of an RNA polymerase binding site at an early T7 promoter. Proc Natl Acad Sci USA 72:784–788
Rosenkrantz MS, Dingman DW, Sonenshein AL (1985) Bacillus subtilis citB gene is regulated synergistically by glucose and glutamine. J Bacteriol 164:155–164
Schmitz A, Galas D (1979) The interaction of RNA polymerase and lac repressor with the lac control region. Nucleic Acids Res 6:111–137
Shen W-F, Squires C, Squires CL (1982) Nucleotide sequence of the rrnG ribosomal RNA promoter region of Escherichia coli. Nucleic Acids Res 10:3303–3313
Stefano J, Gralla J (1982) Spacer mutations in the lac ps promoter. Proc Natl Acad Sci USA 79:1069–1072
Stewart GC, Bott KF (1983) DNA sequence of the tandem ribosomal RNA promoter for B. subtilis operon rrnB. Nucleic Acids Res 11:6289–6300
Travers AA, Lamond AL, Mace HAF, Berman ML (1983) RNA polymerase interactions with the upstream region of the E. coli tyrT promoter. Cell 35:265–273
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Communicated by G.R. Smith
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Le Grice, S.F.J., Shih, CC., Whipple, F. et al. Separation and analysis of the RNA polymerase binding sites of a complex Bacillus subtilis promoter. Molec. Gen. Genet. 204, 229–236 (1986). https://doi.org/10.1007/BF00425503
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DOI: https://doi.org/10.1007/BF00425503