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Molecular analysis and nucleotide sequence of finQ, a transcriptional inhibitor of the F plasmid transfer genes

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Summary

We report the cloning of finQ, a gene coding for fertility inhibition of the F plasmid, from the IncI R factor R820a. The finQ gene was mapped precisely within a 1.24 kb region by ptac-transposase+mini-kan mutagenesis and its product, FinQp, identified as a single polypeptide by means of SDS-polyacrylamide gel electrophoresis. Nucleotide sequencing of the finQ region allowed elucidation of the FinQp amino acid sequence and determination of its precise molecular weight as 39895 Da. Analysis of the predicted amino acid sequence indicated that FinQp is a positively charged protein possessing a helix-turn-helix DNA binding motif. We propose a possible model for the mechanism by which FinQp terminates transcription within the F plasmid tra region. DNA-DNA hybridization established that all FinQ+ R factors examined have an homologous finQ gene.

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References

  • Achtman M, Willetts N, Clark AJ (1971) Beginning a genetic analysis of conjugational transfer determined by the F factor in Escherichia coli by isolation and characterization of transferdeficient mutants. J Bacteriol 106:529–538

    Google Scholar 

  • Achtman M, Manning PA, Kusecek B, Schwuchow S, Willetts N (1980) A genetic analysis of F sex factor cistrons needed for surface exclusion in Escherichia coli. J Mol Biol 138:779–795

    Google Scholar 

  • Casadaban MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:179–207

    Google Scholar 

  • Chang ACY, Cohen SN (1978) Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol 134:1141–1156

    Google Scholar 

  • Cheah K-C, Skurray R (1986) The conjugally-derepressed F plasmid carries an IS3 insertion within finO sequences. J Gen Microbiol 132:3269–3275

    Google Scholar 

  • Cram D, Ray A, O'Gorman L, Skurray R (1984) Transcriptional analysis of the leading region in F plasmid DNA transfer. Plasmid 11:221–233

    Google Scholar 

  • Datta N, Olarte J (1974) R factors in strains of Salmonella typhi and Shigella dysenteriae isolated during epidemics in Mexico: classification and compatibility. Antimicrob Agents Chemother 5:310–317

    Google Scholar 

  • Finnegan DJ, Willetts NS (1971) Two classes of Flac mutants insensitive to transfer inhibition by an F-like R factor. Mol Gen Genet 111:256–264

    Google Scholar 

  • Finnegan D, Willetts N (1973) The site of action of the F transfer inhibitor. Mol Gen Genet 127:307–316

    Google Scholar 

  • Gaffney D, Skurray R, Willetts N (1983) Regulation of the F conjugation genes studied by hybridization and tra-lacZ fusion. J Mol Biol 168:103–122

    Google Scholar 

  • Galas DJ, Eggert M, Waterman MS (1985) Rigorous pattern-recognition methods for DNA sequences: analysis of promoter sequences from Escherichia coli. J Mol Biol 186:117–128

    Google Scholar 

  • Garner J, Osguthorpe DJ, Robson B (1978) Analysis of the accuracy and implications of simple methods for predicting the secondary structure of globular proteins. J Mol Biol 120:97–120

    Google Scholar 

  • Gasson MJ, Willetts NS (1975) Five control systems preventing transfer of the Escherichia coli K12 sex factor F. J Bacteriol 122:518–525

    Google Scholar 

  • Gasson MJ, Willetts NS (1977) Further characterization of the F fertility inhibition systems of “unusual” Fin+ plasmids. J Bacteriol 133:413–420

    Google Scholar 

  • Gouy M, Gautier C (1982) Codon usage in bacteria: correlation with gene expressivity. Nucleic Acids Res 10:7055–7074

    Google Scholar 

  • Grindley JN, Anderson ES (1971) I-like resistance factors with the fi+ character. Genet Res 17:267–271

    Google Scholar 

  • Hedges RW, Datta JN, Coetzee JN, Dennison S (1973) R factors from Proteus morganii. J Gen Microbiol 77:249–259

    Google Scholar 

  • Helmuth R, Achtman M (1975) Operon structure of DNA transfer cistrons of the F sex factor. Nature 257:652–656

    Google Scholar 

  • Ippen-Ihler KA, Minkley Jr EG (1986) The conjugation system of F, the fertility factor of Escherichia coli. Annu Rev Genet 20:593–624

    Google Scholar 

  • Jalajakumari MB, Guidolin A, Buhk HJ, Manning PA, Ham LM, Hodgson ALM, Cheah KC, Skurray RA (1987) Surface exclusion genes traS and traT of the F sex factor of Escherichia coli K-12: determination of the nucleotide sequence and promoter and terminator activities. J Mol Biol 198:1–11

    Google Scholar 

  • Kahn M, Kolter R, Thomas C, Figurski D, Meyer R, Remaut E, Helinski DR (1979) Plasmid cloning vehicles derived from plasmids ColE1, F, R6K, and RK2. Methods Enzymol 68:268–280

    Google Scholar 

  • Low KB (1973) Rapid mapping of conditional and auxotrophic mutants of Escherichia coli K-12. J Bacteriol 113:798–812

    Google Scholar 

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

    Google Scholar 

  • Maxam A, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560

    Google Scholar 

  • Messing J, Vieira J (1982) The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers. Gene 19:259–268

    Google Scholar 

  • Meynell E (1973) Pseudo-fi+ I-like sex factor, R62 (I), selective for increased pilus synthesis. J Bacteriol 113:502–503

    Google Scholar 

  • Miller JF, Lanka E, Malamy MH (1985) F factor inhibition of conjugal transfer of broad-host-range plasmid RP4: requirement for the protein product of pif operon regulatory gene pifC. J Bacteriol 163:1067–1073

    Google Scholar 

  • O'Farrell PH, Kutter E, Nakanishi M (1980) A restriction map of the bacteriophage T4 genome. Mol Gen Genet 179:421–435

    Google Scholar 

  • Oka A, Sugisaki H, Takanami M (1981) Nucleotide sequence of the kanamycin resistance transposon Tn903. J Mol Biol 147:217–226

    Google Scholar 

  • Pabo CO, Sauer RT (1984) Protein-DNA recognition. Annu Rev Biochem 53:293–321

    Google Scholar 

  • Pestka S, Daugherty BL, Jung V, Hotta K, Pestka RK (1984) Anti-mRNA: specific inhibition of translation of single mRNA molecules. Proc Natl Acad Sci USA 81:7525–7528

    Google Scholar 

  • Ray A, Skurray R (1983) Cloning and polypeptide analysis of the leading region in F plasmid DNA transfer. Plasmid 9:262–272

    Google Scholar 

  • Rosenberg M, Court D (1979) Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet 13:319–353

    Google Scholar 

  • Sancar A, Hack AM, Rupp WD (1979) Simple method for identification of plasmid-coded proteins. J Bacteriol 137:692–693

    Google Scholar 

  • Sharp PM, Li W-H (1986) Codon usage in regulatory genes in Escherichia coli does not reflect selection of ‘rare’ condons. Nucleic Acids Res 14:7737–7749

    Google Scholar 

  • Staden R (1986) The current status and portability of our sequence handling software. Nucleic Acids Res 14:217–231

    Google Scholar 

  • Tinoco Jr I, Borer PN, Dengler B, Levine MD, Uhlenbeck OC, Gothers DM, Gralla J (1973) Improved estimation of secondary structure in ribonucleic acids. Nature 246:40–41

    Google Scholar 

  • Way JC, Davis MA, Morisato D, Roberts DE, Kleckner N (1984) New Tn10 derivatives for transposon mutagenesis and for construction of lacZ operon fusions by transposition. Gene 32:369–379

    Google Scholar 

  • Willetts N (1977) The transcriptional control of fertility in F-like plasmids. J Mol Biol 112:141–148

    Google Scholar 

  • Willetts NS, Finnegan DJ (1970) Characterization of an E. coli K12 strain carrying both an F prime and R factor. Genet Res 16:113–122

    Google Scholar 

  • Willetts NS, McIntire S (1978) Isolation and characterization of λtra transducing phages from EDEFL223 (Flac traB::EDλ4). J Mol Biol 126:525–549

    Google Scholar 

  • Willetts NS, Paranchych W (1974) Inhibition of Flac transfer by the Fin+ I-like plasmid R62. J Bacteriol 120:101–105

    Google Scholar 

  • Willetts N, Skurray R (1980) The conjugation system of F-like plasmids. Annu Rev Genet 14:41–76

    Google Scholar 

  • Willetts N, Skurray R (1987) Structure and function of the F factor and mechanism of conjugation. In: Ingraham JL, Low KB, Magasanik B, Neidhart FC, Schaechter M, Umbarger HE (eds) Escherichia coli and Salmonella typhimurium: Cellular and molecular biology. Am Soc Microbiol, Washington DC, pp 1110–1133

    Google Scholar 

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Communicated by N.D.F. Grindley

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Ham, L.M., Skurray, R. Molecular analysis and nucleotide sequence of finQ, a transcriptional inhibitor of the F plasmid transfer genes. Mol Gen Genet 216, 99–105 (1989). https://doi.org/10.1007/BF00332236

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