A genetic dissection of the LlaJI restriction cassette reveals insights on a novel bacteriophage resistance system.

BMC Microbiology
Jonathan O'DriscollDouwe van Sinderen

Abstract

Restriction/modification systems provide the dual function of protecting host DNA against restriction by methylation of appropriate bases within their recognition sequences, and restriction of foreign invading un-methylated DNA, such as promiscuous plasmids or infecting bacteriphage. The plasmid-encoded LlaJI restriction/modification system from Lactococcus lactis recognizes an asymmetric, complementary DNA sequence, consisting of 5'GACGC'3 in one strand and 5'GCGTC'3 in the other and provides a prodigious barrier to bacteriophage infection. LlaJI is comprised of four similarly oriented genes, encoding two 5mC-MTases (M1.LlaJI and M2.LlaJI) and two subunits responsible for restriction activity (R1.LlaJI and R2.LlaJI). Here we employ a detailed genetic analysis of the LlaJI restriction determinants in an attempt to characterize mechanistic features of this unusual hetero-oligomeric endonuclease. Detailed bioinformatics analysis confirmed the presence of a conserved GTP binding and hydrolysis domain within the C-terminal half of the R1.LlaJI amino acid sequence whilst the N-terminal half appeared to be entirely unique. This domain architecture was homologous with that of the "B" subunit of the GTP-dependent, methyl-specific McrBC...Continue Reading

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Citations

Jul 1, 2008·Journal of Biological Dynamics·Nakul ChitnisRichard Steketee
May 29, 2012·Nucleic Acids Research·Kamil SteczkiewiczKrzysztof Ginalski
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Datasets Mentioned

BETA
GM17

Methods Mentioned

BETA
GTPase
GTPases
PCR
electrophoresis
Assay

Software Mentioned

DNASTAR package
seqman
BLAST
DNASTAR

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