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Plasmid RP4 encodes two forms of a DNA primase

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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.

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References

  • Barth PT, Grinter NJ (1977) Map of plasmid RP4 derived by insertion of transposon C. J Mol Biol 113:455–474

    PubMed  Google Scholar 

  • 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 439–448

    Google Scholar 

  • 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 4:121–136

    PubMed  Google Scholar 

  • Boulnois GJ, Wilkins BM (1979) A novel priming system for conjugal synthesis of an IncIα plasmid in recipients. Mol Gen Genet 175:275–279

    PubMed  Google Scholar 

  • Boulnois GJ, Wilkins BM, Lanka E (1982) Overlapping genes at the DNA primase locus of the large plasmid Coll. Nucl Acids Res 10:855–869

    PubMed  Google Scholar 

  • 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 151:1–7

    PubMed  Google Scholar 

  • 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 144:1126–1138

    PubMed  Google Scholar 

  • Danner DB (1982) Recovery of DNA fragments from gels by transfer to DEAE-paper in an electrophoresis chamber. Anal Biochem 125:139–142

    PubMed  Google Scholar 

  • Datta N, Hedges RW, Shaw EJ, Sykes RB, Richmond MH (1971) Properties of an R factor from Pseudomonas aeruginosa. J Bacteriol 108:1244–1249

    PubMed  Google Scholar 

  • Dunn JJ, Studier FW (1983) Complete nucleotide sequence of bacteriophage T7 DNA and the locations of T7 genetic elements. J Mol Biol 166:477–535

    PubMed  Google Scholar 

  • Holmes DS, Quigley M (1981) A rapid boiling method for the preparation of bacterial plasmids. Anal Biochem 114:193–197

    PubMed  Google Scholar 

  • Kröger M, Kröger-Block A (1982) A flexible new computer program for handling DNA sequence data. Nucl Acids Res 10:229–236

    PubMed  Google Scholar 

  • Langridge J, Langridge P, Bergquist PL (1980) Extraction of nucleic acids from agarose gels. Anal Biochem 103:264–271

    PubMed  Google Scholar 

  • Lanka E, Barth PT (1981) Plasmid RP4 specifies a deoxyribonucleic acid primase involved in its conjugal transfer and maintenance. J Bacteriol 148:769–781

    PubMed  Google Scholar 

  • Lanka E, Lurz R, Fürste JP (1983) Molecular cloning and mapping of SphI restriction fragments of plasmid RP4. Plasmid 10:303–307

    PubMed  Google Scholar 

  • Lanka E, Scherzinger E, Günther E, Schuster H (1979) A DNA primase specified by I-like plasmids. Proc Natl Acad Sci USA 76:3632–3636

    PubMed  Google Scholar 

  • Lichtenstein C, Brenner S (1982) Unique insertion site of Tn7 in the E. coli chromosome. Nature 297:601–603

    PubMed  Google Scholar 

  • Linney E, Hayashi M (1973) Two proteins of gene A of ΦX174. Nature New Biol 245:6–8

    PubMed  Google Scholar 

  • Mandel M, Higa A (1970) Calcium-dependent bacteriophage DNA infection. J Mol Biol 53:159–162

    PubMed  Google Scholar 

  • 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 499–560

    Google Scholar 

  • Novick RP, Clowes RC, Cohen SN, Curtiss III R, Datta N, Falkow S (1976) Uniform nomenclature for bacterial plasmids: a proposal. Bacteriol Rev 40:168–189

    PubMed  Google Scholar 

  • Peden KWC (1983) Revised sequence of the tetracycline-resistance gene of pBR322. Gene 22:277–280

    PubMed  Google Scholar 

  • Prentki P, Karch F, Iida S, Meyer J (1981) The plasmid cloning vector pBR325 contains a 482 base-pair-long inverted duplication. Gene 14:289–299

    Article  PubMed  Google Scholar 

  • 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 265:687–695

    PubMed  Google Scholar 

  • 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 272:410–414

    PubMed  Google Scholar 

  • Shaw JE, Murialdo H (1980) Morphogenetic genes C and Nu3 overlap in bacteriophage λ. Nature 283:30–35

    PubMed  Google Scholar 

  • 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 71:1342–1346

    PubMed  Google Scholar 

  • Smith RA, Parkinson JS (1980) Overlapping genes at the cheA locus of Escherichia coli. Proc Natl Acad Sci USA 77:5370–5374

    PubMed  Google Scholar 

  • Stöffler G, Wittmann HG (1971) Sequence differences of Escherichia coli 30S ribosomal proteins as determined by immunochemical methods. Proc Natl Acad Sci USA 68:2283–2287

    PubMed  Google Scholar 

  • Stüber D, Bujard H (1981) Organization of transcriptional signals in plasmids pBR322 and pACYC184. Proc Natl Acad Sci USA 78:167–171

    PubMed  Google Scholar 

  • 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 76:4350–4354

    PubMed  Google Scholar 

  • Wilkins BM, Boulnois GJ, Lanka E (1981) A plasmid DNA primase active in discontinuous bacterial DNA replication. Nature 290:217–221

    PubMed  Google Scholar 

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Communicated by J. Schell

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Lanka, E., Lurz, R., Kröger, M. et al. Plasmid RP4 encodes two forms of a DNA primase. Molec. Gen. Genet. 194, 65–72 (1984). https://doi.org/10.1007/BF00383499

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