Lactococcus lactis phage operon coding for an endonuclease homologous to RuvC

Molecular Microbiology
E BidnenkoM C Chopin

Abstract

The function of the Lactococcus lactis bacteriophage bIL66 middle time-expressed operon (M-operon), involved in sensitivity to the abortive infection mechanism AbiD1, was examined. Expression of the M-operon is detrimental to Escherichia coli cells, induces the SOS response and is lethal to recA and recBC E. coli mutants, which are both deficient in recombinational repair of chromosomal double-stranded breaks (DSBs). The use of an inducible expression system allowed us to demonstrate that the M-operon-encoded proteins generate a limited number of randomly distributed chromosomal DSBs that are substrates for ExoV-mediated DNA degradation. DSBs were also shown to occur upstream of the replication initiation point of unidirectionally theta-replicating plasmids. The characteristics of the DSBs lead us to propose that the endonucleolytic activity of the M-operon is not specific to DNA sequence, but rather to branched DNA structures. Genetic and physical analysis performed with different derivatives of the M-operon indicated that two orfs (orf2 and orf3) are needed for nucleolytic activity. The orf3 product has amino acid homology with the E. coli RuvC Holliday junction resolvase. By site-specific mutagenesis, we have shown that one ...Continue Reading

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Citations

Sep 28, 2002·FEMS Microbiology Letters·Elena BidnenkoJamila Anba
May 20, 2005·Journal of Bacteriology·Louis-Charles FortierSylvain Moineau
Dec 19, 2012·Annual Review of Food Science and Technology·Julie E Samson, Sylvain Moineau
Sep 19, 2015·Standards in Genomic Sciences·Atteyet F YassinNikos C Kyrpides
Feb 20, 2016·Scientific Reports·James MurphyDouwe van Sinderen
Nov 3, 2014·Annual Review of Virology·Ron L DyPeter C Fineran
Feb 22, 2005·Molecular Microbiology·Fiona A CurtisGary J Sharples
Aug 28, 2013·Nature Reviews. Microbiology·Julie E SamsonSylvain Moineau
Mar 30, 2010·Nature Reviews. Microbiology·Simon J LabrieSylvain Moineau
Mar 31, 2015·Applied and Environmental Microbiology·Maxime Bélanger, Sylvain Moineau
Jun 30, 2004·Journal of Molecular Biology·Gary J SharplesEdward L Bolt
Jun 28, 2005·Current Opinion in Microbiology·Marie-Christine ChopinElena Bidnenko

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