Differential distribution of novel restriction-modification systems in clonal lineages of Neisseria meningitidis - PubMed (original) (raw)
Differential distribution of novel restriction-modification systems in clonal lineages of Neisseria meningitidis
H Claus et al. J Bacteriol. 2000 Mar.
Abstract
Using representational difference analysis, we isolated novel meningococcal restriction-modification (R-M) systems. NmeBI, which is a homologue of the R-M system HgaI of Pasteurella volantium, was present in meningococci of the ET-5 complex and of lineage III. NmeAI was found in serogroup A, ET-37 complex, and cluster A4 meningococci. NmeDI was harbored by meningococci of the ET-37 complex and of cluster A4, but not by serogroup A meningococci. Two of the R-M systems, NmeBI and NmeDI, were located at homologous positions between the phenylalanyl-tRNA synthetase genes pheS and pheT, which appeared to be a preferential target for the insertion of foreign DNA in meningococci. The distribution of the three R-M systems was tested with 103 meningococcal strains comprising 49 sequence types. The vast majority of the strains had either NmeBI, NmeAI, or both NmeAI and NmeDI. Using cocultivation experiments, we could demonstrate that NmeBI, which was present in ET-5 complex meningococci, was responsible for a partial restriction of DNA transfer from meningococci of the ET-37 complex to meningococci of the ET-5 complex.
Figures
FIG. 1
(A) Schematic depiction of the insertions between the pheS and pheT genes (encoding the alpha and beta chains of the PheRS) in different meningococcal strains. The length of the insertion in strain Z2491 is 1,803 bp, that in strain MC58 is 3,390 bp, and that in strain 2120 is 3,013 bp. The A+T content is given for the entire insertion and the genes flanking the insertion, respectively. The orientations of the putative ORFs are indicated by arrows. The restriction sites used for the construction of a _Nme_BI deletion mutant of strain MC58 are indicated. (B) Schematic depiction of the location of the _Nme_AI system of strain Z2491 based on the preliminary sequence data released by the meningococcus genome sequencing project (
http://www.sanger.ac.uk./Projects/N-meningitidis/
). The orientations of the putative ORFs are indicated by arrows. The numbers indicate the A+T content of the ORFs and the intergenic regions, respectively.
FIG. 2
Susceptibility of chromosomal DNA to cleavage by _Hga_I. (A) Uncut chromosomal DNA. (B) Chromosomal DNA digested with _Hga_I. (C) Chromosomal DNA digested with both _Hin_dIII and _Eco_RI. Lanes: MC58, wild-type strain (ET-5 complex); MC58Δ, _Nme_BI knockout mutant of MC58; Z2491, N. meningitidis serogroup A (subgroup IV-1); 2120, N. meningitidis serogroup C (ET-37 complex); FA1090, N. gonorrhoeae.
FIG. 3
RNA dot blot hybridizations. A total of 20 μg of RNA of the ET-37 complex strain 2120 and of the ET-5 complex strain MC58, respectively, were dotted onto nylon membranes and hybridized with probes of the MTase genes of the R-M systems _Nme_AI, _Nme_BI, and _Nme_DI. Hybridization with a probe specific to the siaA gene was performed as positive control because the siaA gene is transcribed in both serogroup B and C meningococci.
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