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Suppression of the Escherichia coli dnaA46 mutation by a mutation in trxA, the gene for thioredoxin

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Summary

The dasC mutation, an extragenic suppressor of dnaA46, was mapped by P1 transduction near the rep, trxA, rho region of the Escherichia coli chromosome. The dasC mutation could not be separated from trxA by P1 transduction indicating that dasC and trxA are allelic. Multicopy plasmids containing an intact trxA gene were able to reverse the suppressive effect of the dasC mutation on the dnaA46 mutation. Introduction of a frameshift mutation into the cloned trxA coding region abolished the ability of these recombinant plasmids to reverse the suppressive effect. These results indicate that dasC is allelic with trxA, the gene encoding thioredoxin.

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References

  • Adler S, Modrich P (1985) T7-induced DNA polymerase. Requirement for thioredoxin sulfhydryl groups. J Biol Chem 258:6956–6962

    Google Scholar 

  • Anderson RP, Miller CG, Roth JR (1976) Tandem duplications of the histidine operon observed following generalized transduction in Salmonella typhimurium. J Mol Biol 105:201–218

    Google Scholar 

  • Atlung T (1981) Analysis of seven dnaA suppressor loci in Escherichia coli. ICN-UCLA Symp Mol Cell Biol 22:297–314

    Google Scholar 

  • Bagdasarian MM, Izakowska M, Bagdasarian M (1977) Suppression of the DnaA phenotype by mutations in the rpoB cistron of ribonucleic acid polymerase in Salmonella typhimurium and Escherichia coli. J Bacteriol 130:577–582

    Google Scholar 

  • Benton WD, Davis RW (1977) Screening of λgt recombinant clones by hybridization to single plaques in situ. Science 196:180–182

    Google Scholar 

  • Brown S, Albrechtsen B, Pedersen S, Klemm P (1982) Localization and regulation of the structural gene for transcription-termination factor rho of Escherichia coli. J Mol Biol 162:283–298

    Google Scholar 

  • Buchanan BB, Wolosiuk RA, Schurmann P (1979) Thioredoxin and enzyme regulation. Trends Biochem Sci 4:93–96

    Google Scholar 

  • Chakraborty T, Yoshinaga K, Lother H, Messer W (1982) Purification of the E coli dnaA gene product. EMBO J 1:1545–1549

    Google Scholar 

  • Chamberlin M (1974) Isolation and characterization of prototrophic mutants of Escherichia coli unable to support the intracellular growth of T7. J Virol 14:509–516

    Google Scholar 

  • Clewell DB, Helinski DR (1969) Supercoiled circular DNA-protein complex in Escherichia coli: purification and induced conversion to an open circular DNA form. Proc Natl Acad Sci USA 62:1159–1166

    Google Scholar 

  • Davis RW, Thomas M, Cameron J, St John TP, Scherer S, Padgett RA (1980) Rapid DNA isolations for enzymatic and hybridization analysis. Methods Enzymol 65 (Part 1):404–411

    Google Scholar 

  • de Massey B, Fayet O, Kogoma T (1984) Multiple origin usage for DNA replication in sdrA (rnh) mutants of Escherichia coli K-12. Initiation in the absence of oriC. J Mol Biol 178:227–236

    Google Scholar 

  • Driver RP, Lawther RP (1985) Physical analysis of deletion mutations in the ilvGEDA operon of Escherichia coli K-12. J Bacteriol 162:598–606

    Google Scholar 

  • Eklund H, Cambillau C, Sjoberg B-M, Holmgren A, Jörnvall H, Höög J-O, Branden C-I (1984) Conformational and functional similarities between glutaredoxin and thioredoxins. EMBO J 3:1443–1449

    Google Scholar 

  • Emr SD, Hanley-Way S, Silhavy TJ (1981) Suppressor mutations that restore export of a protein with a defective signal sequence. Cell 23:79–88

    Google Scholar 

  • Filutowicz M, Jonczyk P (1983) The gyrB gene product functions in both initiation and chain polymerization of Escherichia coli chromosome replication. Suppression of the initiation deficiency in gyrB-ts mutants by a class of rpoB mutants. Mol Gen Genet 191:282–287

    Google Scholar 

  • Fuller RS, Kornberg A (1983) Purified dnaA protein in initiation of replication at the Escherichia coli chromosomal origin of replication. Proc Natl Acad Sci USA 80:5817–5821

    Google Scholar 

  • Fuller RS, Funnell BE, Kornberg A (1984) The dnaA protein complex with the E coli chromosomal replication origin (oriC) and other DNA sites. Cell 38:889–900

    Google Scholar 

  • Gray JE, Patin DW, Calhoun DH (1981) Identification of the protein products of the rrnC, ilv, rho region of the Escherichia coli K12 chromosome. Mol Gen Genet 183:428–436

    Google Scholar 

  • Hirota Y, Mordoh J, Jacob F (1970) On the process of cellular division in Escherichia coli. III. Thermosensitive mutants of Escherichia coli altered in the process of DNA initiation. J Mol Biol 53:369–387

    Google Scholar 

  • Holmgren A (1976) Hydrogen donor system for Escherichia coli ribonucleoside-diphosphate reductase dependent upon glutathione. Proc Natl Acad Sci USA 73:2275–2279

    Google Scholar 

  • Höög J-O, von Bahr-Lindstrom H, Josephson S, Wallace BJ, Kushner SR, Jörnvall H, Holmgren A (1984) Nucleotide sequence of the thioredoxin gene from Escherichia coli. Biosci Rep 4:917–923

    Google Scholar 

  • Huber HE, Russel M, Model P, Richardson CC (1986) Interaction of mutant thioredoxins of Escherichia coli with the gene 5 protein of phage T7. J Biol Chem 261:15006–15012

    Google Scholar 

  • Jarvik J, Botstein D (1973) A genetic method for determining the order of events in a biological pathway. Proc Natl Acad Sci USA 70:2046–2050

    Google Scholar 

  • Kaguni J, Ray DS (1979) Cloning of a functional replication origin of phage G4 into the genome of phage M13. J Mol Biol 135:863–878

    Google Scholar 

  • Kogoma T (1976) Two types of temperature sensitivity in DNA replication of an Escherichia coli dnaB mutant. J Mol Biol 103:191–197

    Google Scholar 

  • Lark KG (1972) Evidence for direct involvement of RNA in the initiation of DNA replication in Escherichia coli 15T. J Mol Biol 64:47–60

    Google Scholar 

  • Laurent T, Moore EC, Reichard P (1964) Enzymatic synthesis of deoxyribonucleotides. Isolation and characterization of thioredoxin, the hydrogen donor from Escherichia coli B. J Biol Chem 239:3436–3444

    Google Scholar 

  • Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY

    Google Scholar 

  • Mark DF, Richardson CC (1976) Escherichia coli thioredoxin: A subunit of bacteriophage T7 DNA polymerase. Proc Natl Acad Sci USA 73:780–784

    Google Scholar 

  • Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York

    Google Scholar 

  • Ogawa T, Pickett GG, Kogoma T, Kornberg A (1984) RNase H confers specificity in the dnaA-dependent initiation of replication at the unique origin of the E. coli chromosome in vivo and in vitro. Proc Natl Acad Sci USA 81:1040–1044

    Google Scholar 

  • Ogawa T, Baker TA, van der Ende A, Kornberg A (1985) Initiation of enzymatic replication at the origin of the Escherichia coli chromosome: contributions of RNA polymerase and primase. Proc Natl Acad Sci USA 82:3562–3566

    Google Scholar 

  • Pigiet VP, Schuster BJ (1986) Thioredoxin-catalysed refolding of disulfide-containing proteins. Proc Natl Acad Sci USA 83:7643–7647

    Google Scholar 

  • Pinkham JL, Platt T (1983) The nucleotide sequence of the rho gene of E. coli K-12. Nucleic Acids Res 11:3531–3545

    Google Scholar 

  • Russel M, Model P (1984a) Characterization of the cloned fip gene and its product. J Bacteriol 157:526–532

    Google Scholar 

  • Russel M, Model P (1984b) Replacement of the fip gene of Escherichia coli by an inactive gene cloned on a plasmid. J Bacteriol 159:1034–1039

    Google Scholar 

  • Russel M, Model P (1985) Thioredoxin is required for filamentous phage assembly. Proc Natl Acad Sci USA 82:29–33

    Google Scholar 

  • Russel M, Model P (1986) The role of thioredoxin in filamentous phage assembly. Construction, isolation, and characterization of mutant thioredoxins. J Biol Chem 261:14997–15005

    Google Scholar 

  • Saito H, Miura KI (1963) Preparation of transforming DNA by phenol treatment. Biochim Biophys Acta 72:619–629

    Google Scholar 

  • Sekimizu K, Bramhill D, Kornberg A (1987) ATP activates dnaA protein in initiating replication of plasmids bearing the origin of the E. coli chromosome. Cell 50:259–265

    Google Scholar 

  • Singer BS, Gold L, Shinedling ST, Colkitt M, Hunter LR, Pribnow D, Nelson MA (1981) Analysis in vivo of translational mutants of the rIIB cistron of bacteriophage T4. J Mol Biol 149:405–432

    Google Scholar 

  • Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98:503–517

    Google Scholar 

  • Tessman I, Fassler JS, Bennett DC (1982) Relative map location of the rep and rho genes of Escherichia coli. J Bacteriol 151:1637–1640

    Google Scholar 

  • van der Ende A, Baker TA, Ogawa T, Kornberg A (1985) Initiation of enzymatic replication at the origin of the Escherichia coli chromosome: primase as the sole priming enzyme. Proc Natl Acad Sci USA 79:7639–7643

    Google Scholar 

  • Wechsler JA, Zdzienicka M (1975) Cryolethal suppressors of thermosensitive dnaA mutations. ICN-UCLA Symp Mol Cell Biol 3:624–639

    Google Scholar 

  • Wu TT (1966) A model for three point analysis of random general transduction. Genetics 54:405–410

    Google Scholar 

  • Zyskind JW, Smith DW (1977) Novel Escherichia coli dnaB mutant: direct involvement of the dnaB252 gene product in synthesis of an origin-ribonucleic acid species during initiation of a round of deoxyribonucleic acid replication. J Bacteriol 129:1476–1486

    Google Scholar 

  • Zyskind JW, Deen T, Smith DW (1977) Temporal sequence of events during the initiation process in Escherichia coli deoxyribonucleic acid replication: roles of the dnaA and dnaC gene products and ribonucleic acid polymerase. J Bacteriol 129:1466–1475

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, Michigan State University, 48824, East Lansing, MI, USA

    Theodore R. Hupp & Jon M. Kaguni

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  1. Theodore R. Hupp
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  2. Jon M. Kaguni
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Communicated by N.D.F. Grindley

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Hupp, T.R., Kaguni, J.M. Suppression of the Escherichia coli dnaA46 mutation by a mutation in trxA, the gene for thioredoxin. Mol Gen Genet 213, 471–478 (1988). https://doi.org/10.1007/BF00339618

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  • DOI: https://doi.org/10.1007/BF00339618

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