Summary
Bacteriophage P1 encodes a site-specific recombinase, Cin, which regulates the alternate expression of tail fibre genes by inverting a DNA segment. To define regions of Cin important for the recombination process, we have isolated and characterised 24 different mutations of the cin gene. Most of these mutations affected amino acids that are highly conserved in other related recombinases. Some of these mutants complement each other in vivo. This intragenic complementation could be due to the assembly of heteromers containing both mutant proteins, suggesting that the active enzyme is at least a dimer.
Similar content being viewed by others
References
Abremski K, Hoess R (1984) Bacteriophage P1 site-specific recombination. J Biol Chem 259:1509–1514
Bruist MF, Glasgow AC, Johnson RC, Simon MI (1987a) Fis binding to the recombinational enhancer of the Hin DNA inversion system. Genes Dev 1:762–772
Bruist MF, Horvath SJ, Hood LE, Steitz TA, Simon MI (1987b) Synthesis of a site-specific DNA binding peptide. Science 235:777–780
Carter P, Bedouelle H, Winter G (1985) Improved oligonucleotide site-directed mutagenesis using M13 vectors. Nucleic Acids Res 13:4431–4443
Grindley NDF, Newman BJ, Wiater LA, Falvey EE (1985) In: Simon M, Herskovitz I (eds) Genome Rearrangement, vol 20, UCLA symposium on molecular and cellular biology. Liss, New York, pp 77–92
Haffter P, Bickle TA (1987) Purification and DNA-binding properties of FIS and Cin, two proteins required for the bacteriophage P1 site-specific recombination system, cin. J Mol Biol 198:579–587
Hatfull GF, Salvo JJ, Falvey EE, Rimphanitchayakit V, Grindley NDP (1988) Site-specific recombination by the γδ resolvase. In: Kingsman AJ, Chater KF, Kingsman SM (eds) Transposition. Cambridge University Press, Cambridge, pp 149–181
Hattori M, Sakaki Y (1986) Dideoxy sequencing method using denatured plasmid templates. Anal Biochem 152:232–238
Heffron F, McCarthy BJ, Ohtsubo H, Ohtsubo E (1979) DNA sequence analysis of the transposon Tn3: Three genes and three sites involved in transposition. Cell 18:1153–1163
Heffron F, Kostriken R, Morita C, Parker R (1980) Tn3 encodes a site-specific recombination system: Identification of essential sequences, genes, and the actual site of recombination. Cold Spring Harbor Symp Quant Biol 45:259–268
Hiestand-Nauer R, Iida S (1983) Sequence of the site-specific recombinase gene cin and of its substrates serving in the inversion of the C segment of bacteriophage P1. EMBO J 2:1733–1740
Huber HE, Iida S, Arber W, Bickle TA (1985a) Site-specific DNA inversion is enhanced by a DNA sequence element in cis. Proc Natl Acad Sci USA 82:3776–3780
Huber HE, Iida S, Bickle TA (1985b) Expression of the bacteriophage P1 cin recombinase gene from its own and heterologous promoters. Gene 34:63–72
Iida S (1984) Bacteriophage P1 carries two related sets of genes determining its host range in the invertible C segment of its genome. Virology 134:421–434
Iida S, Meyer J, Kennedy KE, Arber W (1982) A site-specific, conservative recombination system carried by bacteriophage P1. Mapping the recombinase gene cin and the crossover sites cix for the inversion of the C segment. EMBO J 1:1445–1453
Johnson RC, Simon MI (1985) Hin-mediated site-specific recombination requires two 26 bp recombination sites and a 60 bp recombinational enhancer. Cell 41:781–791
Johnson RC, Bruist MF, Glaccum MB, Simon MI (1984) In vitro analysis of Hin-mediated site-specific recombination. Cold Spring Harbor Symp Quant Biol 49:751–760
Kahmann R, Rudt F, Koch C, Mertens G (1985) G inversion in bacteriophage Mu DNA is stimulated by a site within the invertase gene and a host factor. Cell 41:771–780
Kamp D, Chow LT, Broker TR, Kwoh D, Zipser D, Kahmann R (1978) Site-specific recombination in phage Mu. Cold Spring Harbor Symp Quant Biol 43:1159–1167
Kamp D, Kardas E, Ritthaler W, Sandulache R, Schmucker R, Stern B (1984) Comparative analysis of invertible DNA in phage genomes. Cold Spring Harbor Symp Quant Biol 49:301–311
Kanaar R, van de Putte P, Cozzarelli NR (1988) Gin-mediated DNA inversion: Product structure and the mechanism of strand exchange. Proc Natl Acad Sci USA 85:752–756
Klippel A, Mertens G, Patschinsky T, Kahmann R (1988) The DNA invertase Gin of phage Mu: formation of a covalent complex with DNA via a phosphoserine at amino position 9. EMBO J 7:1229–1237
Koch C, Kahmann R (1986) Purification and properties of the Escherichia coli host factor required for inversion of the G segment in bacteriophage Mu. J Biol Chem 261:15673–15678
Koch C, Mertens G, Rudt F, Kahmann R, Kanaar R, Plasterk RHA, van de Putte P, Sandulache R, Kamp D (1987) The invertible G segment. In: Symonds N, Toussiant A, van de Putte P, Howe MM (eds) Phage Mu. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 75–91
Mertens G, Hoffman A, Blöcker H, Frank R, Kahmann R (1984) Gin-mediated site-specific recombination in bacteriophage Mu DNA: overproduction of the protein and inversion in vitro. EMBO J 3:2415–2421
Mertens G, Klippel A, Fuss H, Blöcker H, Framk R, Kahmann R (1988) Site-specific recombination in bacteriophage Mu: characterization of binding sites for the DNA invertase Gin. EMBO J 7:1219–1227
Miller JH (1972) Nitrosoguanidine mutagenesis. In: Experiments in molecular genetics. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York, pp 125–129
Minton NP (1984) Improved plasmid vectors for the isolation of translational lac gene fusions. Gene 31:269–273
Nash HA, Mizuuchi K, Enquist LW, Weisberg RA (1980) Strand exchange in λ integrative recombination: Genetics, biochemistry and models. Cold Spring Harbor Symp Quant Biol 45:417–428
Plasterk RHA, van de Putte P (1984) Genetic switches by DNA inversions in prokaryotes. Biochim Biophys Acta 782:111–119
Plasterk RHA, Brinkman A, van de Putte P (1983) DNA inversions in the chromosome of Escherichia coli and in bacteriophage MU: Relationship to other site-specific recombination systems. Proc Natl Acad Sci USA 80:5355–5358
Reed RR (1981) Transposon-mediated site-specific recombination: A defined in vitro system. Cell 25:713–719
Reed RR, Grindley NDF (1981) Transposon-mediated site-specific recombination in vitro: DNA cleavage and protein-DNA linkage at the recombination site. Cell 25:721–728
Reed RR, Shibuya GI, Steitz JA (1982) Nucleotide sequence of γ δ resolvase gene and demonstration that its gene product acts as a repressor of transcription. Nature 300:381–383
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74:5463–5467
Silverman M, Zieg J, Mandel G, Simon M (1980) Analysis of the functional components of the phase variation system. Cold Spring Harbor Symp Quant Biol 45:17–26
van de Putte P, Plasterk R, Kuijpers A (1984) A Mu gin complementing function and an invertible DNA region in Escherichia coli K12 are situated on the genetic element e14. J Bacteriol 158:517–522
Zieg J, Simon M (1980) Analysis of the nucleotide sequence of an invertible controlling element. Proc Natl Acad Sci USA 77:4196–4200
Author information
Authors and Affiliations
Additional information
Communicated by H. Hennecke
Rights and permissions
About this article
Cite this article
Haffter, P., Pripfl, T. & Bickle, T.A. A mutational analysis of the bacteriophage P1 cin recombinase gene: Intragenic complementation. Mol Gen Genet 215, 245–249 (1989). https://doi.org/10.1007/BF00339724
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00339724