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Isolation and characterization of new copy mutants of plasmid R1, and identification of a polypeptide involved in copy number control

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Summary

Site-specific deletions and insertions in the replication region of plasmid R1 have generated a new class of copy mutants that are present in the cell with 10–15-fold increased copy number. All mutations described inactivate a copy number control gene which is distinct from another cop inc gene that was identified previously (Molin and Nordström 1980). Insertion of the lac operon lacking the normal lac promoter has been used to determine the direction of transcription of this cop gene. The mutants may all be complemented by wild-type plasmid derivatives and are thus recessive. In incompatibility tests with wild-type R1 plasmids, these mutants are indistinguishable from the wild-type plasmid. It therefore seems that this cop function does not play an important role for the incompatibility function. A polypeptide, molecular weight 11,000, has been identified as being the product of this cop gene.

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References

  • Appleyard RK (1954) Segregation of new lysogenic types during growth of a doubly lysogenic strain derived from Escherichia coli K-12. Genetics 39:440–452

    Google Scholar 

  • Beard JP, Connolly JC (1975) Detection of a protein, similar to the sex pilus subunit, in the outer membrane of Escherichia coli cells carrying a derepressed F-like R-factor. J Bacteriol 122:59–65

    Google Scholar 

  • Bergfors S, Burman LG, Eklöf PH, Nordström K, Tärnvik A (1972) Transferable and non-transferable drug resistance in enteric bacteria isolated from urinary specimens in Northern Sweden. Acta Pathol Microbiol Scand [B] 80:511–518

    Google Scholar 

  • Bertani G (1951) Studies on lysogenesis. I The mode of phage liberation by lysogenic Escherichia coli. J Bacteriol 62:293–300

    Google Scholar 

  • Blohm D, Goebel W (1978) Restriction map of the antibiotic resistance plasmid R1drd-19 and its derivatives pKN102 (R1drd-19B2) and R1drd-16 for the enzymes BamHI, HindIII, EcoRI and SalI. Mol Gen Genet 16:119–127

    Google Scholar 

  • Cabello F, Timmis K, Cohen SN (1976) Replication control in a composite plasmid constructed by in vitro linkage of two distinct replicons. Nature 259:285–290

    Google Scholar 

  • Chou J, Lemaux PG, Casadaban MJ, Cohen SN (1979) Transposition protein of Tn3: identification and characterization of an essential repressor-controlled gene product. Nature 282:801–806

    Google Scholar 

  • Clark JD, Maaløe O (1967) DNA replication and the division cycle in Escherichia coli. J Mol Biol 23:99–112

    Google Scholar 

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

    Google Scholar 

  • Dougan G, Sherratt D (1977) The transposon Tn1 as a probe for studying ColE1 structure and function. Mol Gen Genet 151:151–160

    Google Scholar 

  • Dürwald H, Hoffmann-Berling H (1968) Endonuclease I-deficient and ribonuclease I-deficient Escherichia coli mutants. J Mol Biol 34:331–346

    Google Scholar 

  • Frazer AC, Curtiss III R (1975) Production properties and utility of bacterial minicells. Curr Top Microbiol Immunol 69:1–84

    Google Scholar 

  • Goebel W, Bonewald R (1975) Class of small multicopy plasmids originating from the mutant antibiotic resistance factor Rdrd-19B2. J Bacteriol 123:658–665

    Google Scholar 

  • Gustafsson P, Nordström K (1978) Temperature-dependent and amber copy mutants of plasmid R1drd-19 in Escherichia coli. Plasmid 1:134–144

    Google Scholar 

  • Humphreys GO, Willshaw GA, Anderson EA (1975) A simple method for the preparation of large quantities of pure plasmid DNA. Biochim Biophys Acta 383:457–463

    Google Scholar 

  • Kennedy N, Beutin L, Achtman M, Skurray R, Rahmsdorf U, Herrlich P (1977) Conjugation proteins encoded by the F sex factor. Nature (London) 270:580–585

    Google Scholar 

  • Koltek R, Oertel W, Goebel W (1978) Isolation and characterization of the minimal fragment required for autonomous replication of a copy mutant (pKN102) of the antibiotic resistance factor R1. Mol Gen Genet 162:51–58

    Google Scholar 

  • Kolter R, Helinski DR (1979) Regulation of initiation of DNA replication. Ann Rev Genet 13:355–392

    Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685

    Google Scholar 

  • Matsubara K, Takeda Y (1976) Role of the tof gene in the production and perpetuation of the λdv plasmid. Mol Gen Genet 142:226–230

    Google Scholar 

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

    Google Scholar 

  • Molin S, Stougaard P, Uhlin BE, Gustafsson P, Nordström K (1979) Clustering of genes involved in replication, incompatibility and stable maintenance of the resistance plasmid R1. J Bacteriol 138:70–79

    Google Scholar 

  • Molin S, Nordström K (1980) Control of plasmid R` replication: Functions involved in replication, copy number control, incompatibility and switch-off of replication. J Bacteriol 141:111–120

    Google Scholar 

  • Molin S, Diaz R, Uhlin BE, Nordström K (1980) Runaway replication of plasmid R1 is not caused by loss of replication inhibitor activity of the cop gene. J Bacteriol 143:1046–1048

    Google Scholar 

  • Nordström K, Ingram LC, Lundbäck A (1972) Mutations in R factors of Escherichia coli causing an increased number of R-factor copies per chromosome. J Bacteriol 110:562–569

    Google Scholar 

  • Nordström K, Molin S, Aagaard-Hansen H (1980) Partitioning of plasmid R1 in Escherichia coli. I. Kinetics of loss of plasmid derivatives deleted of the par region. Plasmid 4:215–227

    Google Scholar 

  • Nordström K, Molin S, Aagaard-Hansen H (1981) Partitioning of plasmid R1 in Escherichia coli. II. Incompatibility properties of the partitioning system. Plasmid 5: in press

  • 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

    Google Scholar 

  • Oertel W, Kollek R, Beck E, Goebel W (1979) The nucleotide sequence of a DNA fragment from the replication origin of the antibiotic resistance factor R1drd-19. Mol Gen Genet 171:277–285

    Google Scholar 

  • Pritchard RH, Barth PT, Collins J (1969) Control of DNA synthesis in bacteria. Symp Soc Gen Microbiol 19:263–297

    Google Scholar 

  • Shepard HM, Gelfand GH, Polisky B (1979) Analysis of a recessive plasmid copy number mutant: Evidence for negative control of ColE1 replication. Cell 18:267–275

    Google Scholar 

  • Stougaard P, Molin S, Nordström K, Hansen FG (1981) The nucleotide sequence of the replication control region of the resistance plasmid R1drd-19. Mol Gen Genet 181:116–122

    Google Scholar 

  • Uhlin BE, Nordström K (1975) Plasmid incompatibility and control of replication: Copy mutants of the R-factor R1 in Escherichia coli K-12. J Bacteriol 124:641–649

    Google Scholar 

  • Uhlin BE, Nordström K (1978) A runaway-replication mutant of plasmid R1drd-19: Temperature-dependent loss of copy number control. Mol Gen Genet 165:167–179

    Google Scholar 

  • Zubay G (1973) In vitro synthesis of protein in microbial systems. Annu Rev Gen 7:267–287

    Google Scholar 

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Communicated by D. Sherratt

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Molin, S., Stougaard, P., Light, J. et al. Isolation and characterization of new copy mutants of plasmid R1, and identification of a polypeptide involved in copy number control. Mol Gen Genet 181, 123–130 (1981). https://doi.org/10.1007/BF00339015

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  • DOI: https://doi.org/10.1007/BF00339015

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