Advertisement

Molecular and General Genetics MGG

, Volume 194, Issue 1–2, pp 65–72 | Cite as

Plasmid RP4 encodes two forms of a DNA primase

  • Erich Lanka
  • Rudi Lurz
  • Manfred Kröger
  • Jens P. Fürste
Article

Summary

The pri gene locus of the conjugative broad host range plasmid RP4 maps between coordinates 40.3 and 43.5 and encodes two antigenically related forms of a DNA primase with a molecular mass of 118 and 80 kDa (kilodalton). Genesis of these two products has been examined using Pri+-recombinant plasmids. As shown by deletion analysis, the primase polypeptides are two separate translation products which arise from an in-phase overlapping gene arrangement. It is suggested that transcription of a set of RP4 genes including the pri gene starts at a promoter site within the Tra1 region. In vivo, RP4 mutant primase can apparently substitute for Escherichia coli primase as demonstrated by measuring suppression of the dnaG3(ts) mutant.

Keywords

Escherichia Coli Molecular Mass Polypeptide Gene Locus Recombinant Plasmid 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Barth PT, Grinter NJ (1977) Map of plasmid RP4 derived by insertion of transposon C. J Mol Biol 113:455–474PubMedGoogle Scholar
  2. Barth PT, Tobin L, Sharpe GS (1981) Development of broad host-range plasmid vectors. In: Levy SB, Clowes RC, Koenig EL (eds) Molecular biology, pathogenicity, and ecology of bacterial plasmids. Plenum Publ Corp, New York, p 439–448Google Scholar
  3. Bolivar F (1978) Construction and characterization of new cloning vehicles. III. Derivatives of plasmid pBR322 carrying unique EcoRI sites for selection of EcoRI generated recombinant DNA molecules. Gene 4:121–136PubMedGoogle Scholar
  4. Boulnois GJ, Wilkins BM (1979) A novel priming system for conjugal synthesis of an IncIα plasmid in recipients. Mol Gen Genet 175:275–279PubMedGoogle Scholar
  5. Boulnois GJ, Wilkins BM, Lanka E (1982) Overlapping genes at the DNA primase locus of the large plasmid Coll. Nucl Acids Res 10:855–869PubMedGoogle Scholar
  6. Dalrymple BP, Boulnois GJ, Wilkins BM, Orr E, Williams PH (1982) Evidence for two genetically distinct DNA primase activities specified by plasmids of the B and I incompatibility groups. J Bacteriol 151:1–7PubMedGoogle Scholar
  7. Danbara H, Timmis JK, Lurz R, Timmis KN (1980) Plasmid replication functions: Two distinct segments of plasmid R1, RepA and RepD, express incompatibility and are capable of autonomous replication. J Bacteriol 144:1126–1138PubMedGoogle Scholar
  8. Danner DB (1982) Recovery of DNA fragments from gels by transfer to DEAE-paper in an electrophoresis chamber. Anal Biochem 125:139–142PubMedGoogle Scholar
  9. Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of an R factor from Pseudomonas aeruginosa. J Bacteriol 108:1244–1249PubMedGoogle Scholar
  10. Dunn JJ, Studier FW (1983) Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol 166:477–535PubMedGoogle Scholar
  11. Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197PubMedGoogle Scholar
  12. Kröger M, Kröger-Block A (1982) A flexible new computer program for handling DNA sequence data. Nucl Acids Res 10:229–236PubMedGoogle Scholar
  13. Langridge J, Langridge P, Bergquist PL (1980) Extraction of nucleic acids from agarose gels. Anal Biochem 103:264–271PubMedGoogle Scholar
  14. Lanka E, Barth PT (1981) Plasmid RP4 specifies a deoxyribonucleic acid primase involved in its conjugal transfer and maintenance. J Bacteriol 148:769–781PubMedGoogle Scholar
  15. Lanka E, Lurz R, Fürste JP (1983) Molecular cloning and mapping of SphI restriction fragments of plasmid RP4. Plasmid 10:303–307PubMedGoogle Scholar
  16. Lanka E, Scherzinger E, Günther E, Schuster H (1979) A DNA primase specified by I-like plasmids. Proc Natl Acad Sci USA 76:3632–3636PubMedGoogle Scholar
  17. Lichtenstein C, Brenner S (1982) Unique insertion site of Tn7 in the E. coli chromosome. Nature 297:601–603PubMedGoogle Scholar
  18. Linney E, Hayashi M (1973) Two proteins of gene A of ΦX174. Nature New Biol 245:6–8PubMedGoogle Scholar
  19. Mandel M, Higa A (1970) Calcium-dependent bacteriophage DNA infection. J Mol Biol 53:159–162PubMedGoogle Scholar
  20. 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, p 499–560Google Scholar
  21. Novick RP, Clowes RC, Cohen SN, Curtiss III R, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev 40:168–189PubMedGoogle Scholar
  22. Peden KWC (1983) Revised sequence of the tetracycline-resistance gene of pBR322. Gene 22:277–280PubMedGoogle Scholar
  23. Prentki P, Karch F, Iida S, Meyer J (1981) The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene 14:289–299CrossRefPubMedGoogle Scholar
  24. Sanger F, Air GM, Barrell BG, Brown NL, Coulson AR, Fiddes JC, Hutchison III CA, Slocombe PM, Smith M (1977) Nucleotide sequence of bacteriophage ΦX174 DNA. Nature 265:687–695PubMedGoogle Scholar
  25. Schwarz E, Scherer G, Hobom G, Kössel H (1978) Nucleotide sequence of cro, cII and part of the O gene in phage λ DNA. Nature 272:410–414PubMedGoogle Scholar
  26. Shaw JE, Murialdo H (1980) Morphogenetic genes C and Nu3 overlap in bacteriophage λ. Nature 283:30–35PubMedGoogle Scholar
  27. Shine J, Dalgarno L (1974) The 3′-terminal sequence of Escherichia coli 16S ribosomal RNA: Complementarity to nonsense triplets and ribosome binding sites. Proc Natl Acad Sci USA 71:1342–1346PubMedGoogle Scholar
  28. Smith RA, Parkinson JS (1980) Overlapping genes at the cheA locus of Escherichia coli. Proc Natl Acad Sci USA 77:5370–5374PubMedGoogle Scholar
  29. Stöffler G, Wittmann HG (1971) Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc Natl Acad Sci USA 68:2283–2287PubMedGoogle Scholar
  30. Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171PubMedGoogle Scholar
  31. Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354PubMedGoogle Scholar
  32. Wilkins BM, Boulnois GJ, Lanka E (1981) A plasmid DNA primase active in discontinuous bacterial DNA replication. Nature 290:217–221PubMedGoogle Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • Erich Lanka
    • 1
  • Rudi Lurz
    • 1
  • Manfred Kröger
    • 2
  • Jens P. Fürste
    • 1
  1. 1.Abt. SchusterMax-Planck-Institut für Molekulare GenetikBerlin 33Federal Republic of Germany
  2. 2.Institut für Biologie IIIUniversität FreiburgFreiburgFederal Republic of Germany

Personalised recommendations