The sre gene (ORF469) encodes a site-specific recombinase responsible for integration of the R4 phage genome

The sre gene (ORF469) of the R4 phage encodes a protein similar to the resolvase-DNA invertase family proteins. Insertional gene disruption of sre prevented a lysogen from entering the lytic cycle, implying that Sre protein is a site-specific recombinase needed for excision of the R4 prophage genome (M. Matsuura, T. Noguchi, T. Aida, M. Asayama, H. Takahashi, and M. Shirai, J. Gen. Appl. Microbiol. 41:53-61, 1995). To determine whether this sre gene is also necessary for the integration reaction, we studied its function by integration plasmid analysis. When deletions, frameshifts, and site-directed mutations that caused an amino acid substitution of Ser-17 for Ala were introduced into the sre structural gene, transformation efficiency of Streptomyces parvulus 2297 with these plasmid DNAs was severely reduced. However, an adenine insertion just before the possible initiation codon of the sre gene did not significantly decrease the efficiency. These data suggest that the Sre protein is a site-specific recombinase responsible for integration of the R4 phage genome.

[1]  N. Grindley,et al.  Analysis of gamma delta resolvase mutants in vitro: evidence for an interaction between serine-10 of resolvase and site I of res. , 1986, Proceedings of the National Academy of Sciences of the United States of America.

[2]  D. Hopwood Genetic manipulation in Streptomyces , 1983 .

[3]  T. Isogai,et al.  Construction of phage vectors in Streptomyces: Introduction of the thiostreptone resistant (tsr) gene into R4 phage. , 1984 .

[4]  T. Kunkel Rapid and efficient site-specific mutagenesis without phenotypic selection. , 1985, Proceedings of the National Academy of Sciences of the United States of America.

[5]  J. W. Golden,et al.  Anabaena xisF gene encodes a developmentally regulated site-specific recombinase. , 1994, Genes & development.

[6]  T. Isogai,et al.  High-frequency Protoplast-transfection of Streptomyces parvulus 2297 with Actinophage R4 DNA , 1980 .

[7]  M. Goodfellow,et al.  Actinomycetes in biotechnology , 1988 .

[8]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[9]  M. Asayama,et al.  A GENE ESSENTIAL FOR THE SITE-SPECIFIC EXCISION OF ACTINOPHAGE R4 PROPHAGE GENOME FROM THE CHROMOSOME OF A LYSOGEN , 1995 .

[10]  H. Nash,et al.  Nicking-closing activity associated with bacteriophage lambda int gene product. , 1979, Proceedings of the National Academy of Sciences of the United States of America.

[11]  J. Murray,et al.  Site-specific recombinases: tools for genome engineering. , 1993, Trends in genetics : TIG.

[12]  H. Nash,et al.  Purification and properties of the Escherichia coli protein factor required for lambda integrative recombination. , 1981, The Journal of biological chemistry.

[13]  T. Sato,et al.  The cisA cistron of Bacillus subtilis sporulation gene spoIVC encodes a protein homologous to a site-specific recombinase , 1990, Journal of bacteriology.

[14]  H. Takahashi,et al.  Delimitation of cohesive ends site (cos) of Streptomyces temperate bacteriophage R4. , 1992, Biochemical and biophysical research communications.

[15]  S. Gottesman,et al.  Purification of the bacteriophage lambda xis gene product required for lambda excisive recombination. , 1982, The Journal of biological chemistry.