Cloning and expression of the BamHI restriction-modification system in Bacillus subtilis

Kawakami, Bunsei; Katsuragi, Nobuhiro; Maekawa, Yoichi; Imanaka, Tadayuki

doi:10.1016/0922-338x(90)90050-7

Cited by 2 publications

(1 citation statement)

References 13 publications

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“…An Sph I– Ssp I fragment carrying spc from pBEST517 (Toda et al ., 1996) was used to replace the Sph I– Ssp I fragment of pBR322 (= pYAE7). A Hin dIII– Bgl II fragment carrying Bam HI methyltransferase, control element and endonuclease genes (m c r) from pBamHIRM22 (Kawakami et al ., 1990) (provided by B. Kawakami) and the Eco RI– Bam HI fragment carrying neo from pBEST502 (Itaya et al ., 1989) were used to replace the Eco RI– Bam HI region of pBR322 (= pYAE5). A Bgl II linker (CAGATCTG) was then inserted into the Bal I site of its r gene (= pYAE6).…”

Section: Methodsmentioning

confidence: 99%

Multiplication of a restriction–modification gene complex

Sadykov

Asami

Niki

et al. 2003

Molecular Microbiology

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SummaryPrevious works have suggested that some gene complexes encoding a restriction (R) enzyme and a cognate modification (M) enzyme may behave as selfish mobile genetic elements. RM gene complexes, which destroy 'non-self' elements marked by the absence of proper methylation, are often associated with mobile genetic elements and are involved in various genome rearrangements. Here, we found amplification of a restriction-modification gene complex. Bam HI gene complex inserted into the Bacillus chromosome showed resistance to replacement by a homologous stretch of DNA. Some cells became transformed with the donor without losing Bam HI. In most of these transformants, multiple copies of Bam HI and the donor allele were arranged as tandem repeats. When a clone carrying one copy of each allele was propagated, extensive amplification of Bam HI and the donor unit was observed in a manner dependent on restriction enzyme gene. This suggests that restriction cutting of the genome participates in the amplification. Visualization by fluorescent in situ hybridization revealed that the amplification occurred in single cells in a burst-like fashion that is reminiscent of induction of provirus replication. The multiplication ability in a bacterium with natural capacity for DNA release, uptake and transformation will be discussed in relation to spreading of RM gene complexes.

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Section: Methodsmentioning

confidence: 99%

Multiplication of a restriction–modification gene complex

Sadykov

Asami

Niki

et al. 2003

Molecular Microbiology

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Restriction-modification gene complexes as selfish gene entities: Roles of a regulatory system in their establishment, maintenance, and apoptotic mutual exclusion

Nakayama

Kobayashi

1998

Proc. Natl. Acad. Sci. U.S.A.

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We have reported some type II restrictionmodification (RM) gene complexes on plasmids resist displacement by an incompatible plasmid through postsegregational host killing. Such selfish behavior may have contributed to the spread and maintenance of RM systems. Here we analyze the role of regulatory genes (C), often found linked to RM gene complexes, in their interaction with the host and the other RM gene complexes. We identified the C gene of EcoRV as a positive regulator of restriction. A C mutation eliminated postsegregational killing by EcoRV. The C system has been proposed to allow establishment of RM systems in new hosts by delaying the appearance of restriction activity. Consistent with this proposal, bacteria preexpressing ecoRVC were transformed at a reduced efficiency by plasmids carrying the EcoRV RM gene complex. Cells carrying the BamHI RM gene complex were transformed at a reduced efficiency by a plasmid carrying a PvuII RM gene complex, which shares the same C specificity. The reduction most likely was caused by chromosome cleavage at unmodified PvuII sites by prematurely expressed PvuII restriction enzyme. Therefore, association of the C genes of the same specificity with RM gene complexes of different sequence specificities can confer on a resident RM gene complex the capacity to abort establishment of a second, incoming RM gene complex. This phenomenon, termed ''apoptotic mutual exclusion,'' is reminiscent of suicidal defense against virus infection programmed by other selfish elements. pvuIIC and bamHIC genes define one incompatibility group of exclusion whereas ecoRVC gene defines another.

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Cloning and expression of the BamHI restriction-modification system in Bacillus subtilis

Cited by 2 publications

References 13 publications

Multiplication of a restriction–modification gene complex

Multiplication of a restriction–modification gene complex

Restriction-modification gene complexes as selfish gene entities: Roles of a regulatory system in their establishment, maintenance, and apoptotic mutual exclusion

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