Summary
Gene product 33 of phage T4 is known to be essential in late transcription. Upstream from gene 33 and overlapping its 5′ terminal sequence by 20 bp, we identified an open reading frame coding for a binding protein for double-stranded DNA (DsbA). Gene product DsbA is composed of 89 amino acid residues with a Mr of 10 376 kDa. We purified this protein to homogeneity from over-expressing cells. Gel retardation assays reveal that it binds to DNA and footprint analyses disclose that it interacts preferentially with T4 late promoter regions. At the sites of binding the protein introduces nicks in double-stranded DNA. In vitro transcription assays performed with T4 late modified RNA polymerase on restriction fragments harbouring a T4 late promoter region prove that gene product DsbA enhances transcription from these promoter regions in the presence of gene product 33. Gene dsbA is distinct from gene das which maps close to this genomic region.
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References
Alberts B, Herrick G (1971) DNA cellulose chromatography. Methods Enzymol 21:198–217
Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K12. Genetics 39:440–452
Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA, Struhl K (1989) Current protocols in molecular biology. Greene Publishing Associates and Wiley-Interscience, NY
Biggin MD, Gibson TJ, Hong GF (1983) Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci USA 80:3963–3965
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res 7:1513–1523
Burgess RR (1969) A new method for the large scale purification of E. coli deoxyribonucleic acid-dependent ribonucleic acid polymerase. J Biol Chem 244:6160–6167
Chen EY, Seeburg PH (1985) Supercoiling sequencing: a fast and simple method for sequencing plasmid DNA. DNA 4:165–170
Christensen AC, Young ET (1983) Characterization of T4 transcripts. In: Mathews CK, Kutter EM, Mosig G, Berget PB (eds) Bacteriophage T4. American Society for Microbiology, Washington, DC, pp 184–188
Drivdahl RH, Kutter EM (1990) Inhibition of transcription of cytosine-containing DNA in vitro by the alc gene product of bacteriophage T4. J Bacteriol 172:2716–2727
Elliot T, Geiduschek EP (1984) Defining a bacteriophage T4 late promoter: absence of a “−35” region. Cell 36:211–219
Geiduschek EP, Elliot T, Kassavetis G (1983) Regulation of late gene expression. In: Mathews CK, Kutter EM, Mosig G, Berget PB (eds) Bacteriophage T4. American Society for Microbiology, Washington, DC, pp 189–192
Gram H, Rüger W (1985) Genes 55, αgt, 47 and 46 of Bacteriophage T4: the genomic organization as deduced by sequence analysis. EMBO J 4:257–264
Gribskov M, Burgess RR (1986) σ-factors from E. coli, B. subtilis, phage SPO1 and phage T4 are homologous proteins. Nucleic Acids Res 14:6745–6763
Hahn S, Rüger W (1989) Organization of the bacteriophage T4 genome between map positions 150.745 and 145.824. Nucleic Acids Res 17:6729
Hahn S, Kruse U, Rüger W (1986) The region of phage T4 genes 34, 33 and 59: primary structures and organization on the genome. Nucleic Acids Res 14:9311–9327
Hanahan D (1983) Studies on transformation of Escherichia coli with plasmids. J Mol Biol 166:557–580
Helmann JD, Chamberlin MJ (1988) Structure and function of bacterial σ-factors. Annu Rev Biochem 57:839–872
Hercules K, Wiberg JS (1971) Specific suppression of mutations in genes 46 and 47 by das, a new class of mutations in Bacteriophage T4D. J Virol 8:603–612
Herendeen DR, Kassavetis GA, Barry J, Alberts B, Geiduschek EP (1989) Enhancement of bacteriophage T4 late transcription by components of the T4 DNA replication apparatus. Science 245:952–958
Herendeen DR, Williams KP, Kassavetis GA, Geiduschek EP (1990) An RNA polymerase-binding protein that is required for communication between an enhancer and a promoter. Science 248:573–578
Kassavetis GA, Geiduschek EP (1984) Defining a bacteriophage T4 late promoter: bacteriophage T4 gene 55 protein suffices for directing late promoter recognition. Proc Natl Acad Sci USA 81:5101–5105
Korneluk RG, Quan F, Gravel RA (1985) Rapid and reliable dideoxy sequencing of double-stranded DNA. Gene 40:317–323
Kricker MC, Tindall KR (1989) Direct sequencing of Bacteriophage T4 DNA with a thermostable DNA polymerase. Gene 85:199–204
Kutter EM, Wiberg JS (1968) Degradation of cytosine-containing bacterial and bacteriophage DNA after infection of E. coli B with bacteriophage T4D, wild type and with mutants defective in genes 46, 47 and 56. J Mol Biol 38:395–406
Kutter E, Guttmann B, Mosig G, Rüger W (1990) The Bacteriophage T4 Genomic Map. In: SJ O'Brien (ed) Genetic maps, vol 5. Cold Spring Harbor Laboratory, New York
Liebig HD, Rüger W (1989) Bacteriophage T4 early promoter regions: consensus seqeunces of promoters and ribosome-binding sites. J Mol Biol 208:517–536
Malik S, Goldfarb A (1984) The effect of a bacteriophage T4-induced polypeptide on host RNA polymerase interaction with promoters. J Biol Chem 259:13292–13297
Malik S, Dimitrov M, Goldfarb A (1985) Initiation of transcription by bacteriophage T4-modified RNA polymerase independently of host σ-factor. J Mol Biol 185:83–91
Maxam AM, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Messing J, Crea R, Seeburg P (1981) A system for shotgun DNA sequencing. Nucleic Acids Res 9:309–321
Mickelson C, Wiberg JS (1981) Membrane-associated DNase activity controlled by genes 46 and 47 of bacteriophage T4D and elevated DNase activity associated with the T4 das mutation. J Virol 40:65–77
Nomura M, Matsubara K, Okamoto K, Fujimnura R (1962) Inhibition of host nucleic acid and protein synthesis by bacteriophage T4: its relation to the physical and functional integrity of host chromosome. J Mol Biol 5:535–549
Quirk S, Bell-Pedersen D, Belfort M (1989) Intron mobility in the T-even phages: High frequency inheritance of group I introns promoted by intron open reading frames. Cell 56:455–465
Rabussay D (1982) Bacteriophage T4 infection mechanisms. In: Cohen P, von Heyningen S (eds) Molecular action of toxins and viruses. Elsevier Biochemical Press, New York, pp 219–331
Rabussay D (1983) Phage-evoked changes in RNA polymerase. In: Mathews CK, Kutter EM, Mosig G, Berget PB (eds) Bacteriophage T4. American Society for Microbiology, Washington, DC, pp 167–173
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Schmitz A, Galas DJ (1979) The interaction of RNA polymerase and lac repressor with the lac control region. Nucleic Acids Res 6:111–137
Schneider TD, Stormo GD, Gold L, Ehrenfeucht A (1986) Information content of binding sites on nucleotide sequences. J Mol Biol 188:415–431
Spicer EK, Noble JA, Nossal NG, Konigsberg WH, Williams KR (1982) Bacteriophage T4 gene 45: sequences of the structural gene and its product. Proc Natl Acad Sci USA 257:8972–8979
Stevens A (1972) New small polypeptides associated with DNA-dependent RNA polymerase of Escherichia coli after infection with bacteriophage T4. Proc Natl Sci USA 69:603–604
Stevens A (1974) Desoxyribonucleic acid dependent ribonucleic acid polymerases from two T4 phage-infected systems. Biochemistry 13:493–503
Stevens A, Rhoton JC (1975) Characterization of an inhibitor causing potassium chloride sensitivity of an RNA polymerase from T4 phage-infected Escherichia coli. Biochemistry 14:5074–5079
Strauss F, Varshavsky A (1984) A protein binds to a satellite DNA repeat at three specific sites that would be brought into mutual proximity by DNA folding in the nucleosome. Cell 37:889–901
Tabor S, Richardson CC (1985) A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA 82:1074–1078
Tabor S, Richardson CC (1987) DNA sequencing analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci USA 84:4767–4771
Thomas CA, Abelson J (1960) The isolation and characterization of DNA from bacteriophage. In: Cantoni GL, Davies DR (eds) Procedures in Nucleic Acid Research. Harper and Row, New York, pp 553–561
Tomaschewski J, Rüger W (1987) Nucleotide sequence and primary structures of gene products coded for by the T4 genome between map positions 48.266 kb and 39.166 kb. Nucleic Acids Res 15:3632–3633
Williams JG, Gratzer WB (1971) Limitations of the detergent-polyacrylamide gel electrophoresis method for molecular weight determination of protein. J Chromatogr 57:121–125
Williams KP, Kassavetis GA, Esch FS, Geiduschek EP (1987) Identification of the gene encoding an RNA polymerase binding protein of bacteriophage T4. J Virol 61:597–599
Williams PW, Müller R, Rüger W, Geiduschek EP (1989) Overproduced bacteriophage T4 gene 33 protein binds RNA polymerase. J Bacteriol 171:3579–3582
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Communicated by J.W. Lengeler
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Gansz, A., Kruse, U. & Rüger, W. Gene product dsbA of bacteriophage T4 binds to late promoters and enhances late transcription. Molec. Gen. Genet. 225, 427–434 (1991). https://doi.org/10.1007/BF00261683
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DOI: https://doi.org/10.1007/BF00261683