Skip to main content
Log in

A T7 amber mutant defective in DNA-Binding protein

  • Published:
Molecular and General Genetics MGG Aims and scope Submit manuscript

Summary

A T7 amber mutant, UP-2, in the gene for T7 DNA-binding protein was isolated from mutants that could not grow on sup + ssb-1 bacteria but could grow on glnU ssb-1 and sup + ssb +bacteria. The mutant phage synthesized a smaller amber polypeptide (28,000 daltons) than T7 wild-type DNA-dinding protein (32,000 daltons). DNA synthesis of the UP-2 mutant in sup + ssb-1 cells was severely inhibited and the first round of replication was found to be repressed. The abilities for genetic recombination and DNA repair were also low even in permissive hosts compared with those of wild-type phage. Moreover, recombination intermediate T7 DNA molecules were not formed in UP-2 infected nonpermissive cells. The gene that codes for DNA-binding protein is referred to as gene 2.5 since the mutation was mapped between gene 2 and gene 3.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

References

  • Araki H, Ogawa H (1981) The participation of T7 DNA-binding protein in T7 genetic recombination. Virology 111:509–515

    Google Scholar 

  • Baas PD, Keegstra W, Teertstra WR, Jansg HS (1978) R-loops in bacteriophage ϕX174 RF DNA. J Mol Biol 125:187–205

    Google Scholar 

  • Campbell A (1965) The steric effect in lysogenization by bacteriophage lambda. I. Lysogenization of a partially diploid strain of Escherichia coli K12. Virology 27:329–339

    Google Scholar 

  • Clark AJ, Chamberlin M, Boyce RP, Howard-Flanders P (1966) Abnormal metabolic response to ultraviolet light of a recombination deficient mutant of Escherichia coli K12. J Mol Biol 19:442–454

    Google Scholar 

  • Clowes RC, Hayes W (1968) Experiments in microbial genetics. Wiley, New York

    Google Scholar 

  • Glassberg J, Meyer RR, Kornberg A (1979) Mutant single-strand binding protein of Escherichia coli: genetic and physiological characterization. J Bacteriol 140:14–19

    Google Scholar 

  • Hausmann R (1976) Bacteriophage T7 genetics. Curr Top Microbiol Immunol 75:77–110

    Google Scholar 

  • Ikeda H, Tomizawa J (1965) Transducing fragments in generalized transduction by phage P1. I. Molecular origin of the fragments. J Mol Biol 14:85–109

    Google Scholar 

  • Mark FD, Chase JM, Richardson CC (1977) Genetic mapping of trxA, a gene affecting thioredoxin in Escherichia coli K12. Mol Gen Genet 155:145–152

    Google Scholar 

  • Masukata H, Ogawa T, Ogawa H (1979) The R-loop structure of λ DNA in dnaG mutant of E. coli. Abstracts of the annual meeting of the American Society for Microbiology-1979, 137

  • Meyer RR, Glassberg J, Kornberg A (1979) An Escherichia coli mutant defective in single-strand binding protein is defective in DNA replication. Proc Natl Acad Sci USA 76:1702–1705

    Google Scholar 

  • North R, Molineux IJ (1980) A novel mutant of bacteriophage T7 that is defective in early phage DNA synthesis. Mol Gen Genet 179:683–691

    Google Scholar 

  • Ogawa T (1975) Analysis of the dnaB function of E. coli K12 and the dnaB-like function of P1 prophage. J Mol Biol 94:327–340

    Google Scholar 

  • Ogawa T, Tomizawa J (1967) Abortive lysogenization of bacteriophase lambda b2 and residual immunity of non-lysogenic segregants. J Mol Biol 23:225–245

    Google Scholar 

  • Reuben CR, Gefter ML (1973) A DNA-binding protein induced by bacteriophage T7. Proc Natl Acad Sci USA 70:1846–1850

    Google Scholar 

  • Reuben CR, Gefter ML (1974) A deoxyribonucleic acid-binding protein induced by bacteriophage T7. Purification and properties of the protein. J Biol Chem 249:3843–3850

    Google Scholar 

  • Richardson CC, Romano LJ, Kolodner R, LeClerc JE, Tamanoi F, Engler MJ, Dean FB, Richardson DS (1978) Replication of bacteriophage T7 DNA by purified proteins. Cold Spring Harbor Symp Quant Biol 43:427–440

    Google Scholar 

  • Sancar A, Rupp WD (1979) Cloning of uvrA, lexC and ssb genes of Escherichia coli. Biochem Biophys Res Commun 90:123–129

    Google Scholar 

  • Scherzinger E, Klotz G (1975) Studies of bacteriophage T7 DNA synthesis in vitro. II. Reconstitution of the T7 replication system using purified proteins. Mol Gen Genet 141:233–249

    Google Scholar 

  • Scherzinger E, Litfin F, Jost E (1973) Stimulation of T7 DNA polymerase by a new phage-coded protein. Mol Gen Genet 123:247–262

    Google Scholar 

  • Sevastopoulous CG, Wehr CT, Glaser DA (1977) Large-scale automated isolation of Escherichia coli mutants with thermosensitive DNA replication. Proc Natl Acad Sci USA 74:3485–3489

    Google Scholar 

  • Shapiro AL, Vinuela E, Maizel JV (1967) Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels. Biochem Biophys Res Commun 28:815–820

    Google Scholar 

  • Studier FW (1969) The genetics and physiology of bacteriophage T7. Virology 39:562–574

    Google Scholar 

  • Studier FW (1973) Analysis of bacteriophage T7 early RNAs and proteins on slab gels. J Mol Biol 79:237–248

    Google Scholar 

  • Tomizawa J, Anraku N, Iwama Y (1966) Molecular mechanism of genetic recombination of bacteriophage. VI. A mutant defective in the joining of DNA molecules. J Mol Biol 21:247–253

    Google Scholar 

  • Tsujimoto Y, Ogawa H (1977) Intermediates in genetic recombination of bacteriophage T7 DNA. J Mol Biol 109:423–436

    Google Scholar 

  • Tsujimoto Y, Ogawa H (1978) Intermediates in genetic recombination of bacteriophage T7 DNA. Biological activity and the roles of gene 3 and gene 5. J Mol Biol 125:255–273

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Additional information

Communicated by T. Yura

Rights and permissions

Reprints and permissions

About this article

Cite this article

Araki, H., Ogawa, H. A T7 amber mutant defective in DNA-Binding protein. Molec. Gen. Genet. 183, 66–73 (1981). https://doi.org/10.1007/BF00270140

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00270140

Keywords

Navigation