Chromosomal loci of genes controlling site-specific restriction endonucleases of Bacillus subtilis

Ikawa, Shiro; Shibata, Takehiko; Matsumoto, Kouji; Iijima, Tadako; Saito, Hiuga; Ando, Tetsu

doi:10.1007/bf00270129

Cited by 24 publications

(8 citation statements)

References 36 publications

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“…On the other hand, five restriction-modification systems have been found in B. subtilis Marburg-related bacteria (32). These systems have been introduced into the B. subtilis Marburg chromosome by transformation (10,11,12,33). They are BsuB, BsuC, BsuE, BsuF, and BsuR, and BsuC, BsuF, and BsuR are allelic.…”

Section: Discussionmentioning

confidence: 99%

Molecular Organization of Intrinsic Restriction and Modification Genes Bsu M of Bacillus subtilis Marburg

et al. 2002

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Transcriptional analysis and disruption of five open reading frames (ORFs), ydiO, ydiP, ydiR, ydiS, and ydjA, in the prophage 3 region of the chromosome of Bacillus subtilis Marburg revealed that they are component genes of the intrinsic BsuM restriction and modification system of this organism. The classical mutant strain RM125, which lacks the restriction and modification system of B. subtilis Marburg, lacks the prophage 3 region carrying these five ORFs. These ORFs constitute two operons, the ydiO-ydiP operon and the ydiR-ydiS-ydjA operon, both of which are expressed during the logarithmic phase of growth. The predicted gene products YdiO and YdiP are the orthologues of cytosine DNA methyltransferases. The predicted YdiS product is an orthologue of restriction nucleases, while the predicted YdiR and YdjA products have no apparent paralogues and orthologues whose functions are known. Disruption of the ydiR-ydiS-ydjA operon resulted in enhanced transformation by plasmid DNA carrying multiple BsuM target sequences. Disruption of ydiO or ydiP function requires disruption of at least one of the following genes on the chromosome: ydiR, ydiS, and ydjA. The degrees of methylation of the BsuM target sequences on chromosomal DNAs were estimated indirectly by determining the susceptibility to digestion with XhoI (an isoschizomer of BsuM) of DNAs extracted from the disruptant strains. Six XhoI (BsuM) sites were examined. XhoI digested at the XhoI sites in the DNAs from disruptants with disruptions in both operons, while XhoI did not digest at the XhoI sites in the DNAs from the wild-type strain or from the disruptants with disruptions in the ydiR-ydiS-ydjA operon. Therefore, the ydiO-ydiP operon and the ydiR-ydiS-ydjA operon are considered operons that are responsible for BsuM modification and BsuM restriction, respectively.The existence of an inherent BsuM restriction and modification system in Bacillus subtilis Marburg 168 was first suggested on the basis of the results of an experiment performed with phage 105 and B. subtilis and Bacillus amyloliquefaciens host strains (28). The hsrM1 and nonB mutations (24, 36) which made host cells permissive to phage infection were isolated and mapped at around 50°on the B. subtilis chromosome (24), and it was thought that these mutations were mutations of the endonuclease gene of BsuM. On the other hand, strain RM125 (34) was constructed by transformation of wild-type B. subtilis Marburg 168 (YS11) with DNA from the related strain B. subtilis 202-5 (ϭ IAM1169, a B. amyloliquefaciens strain), which lacked restriction activity against phage 105. Strain RM125 constructed in this way turned out to be deficient in modification activity, as well as restriction activity. However, it was not known whether RM125 lost the same BsuM restriction activity as nonB and hsrM1 strains.The target sequence of the BsuM restriction and modification system of B. subtilis was first predicted to be PyTCGAPu (7, 13), but in another study researchers determined that the BsuM target sequence was CTCGAG by p...

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Section: Discussionmentioning

confidence: 99%

Molecular Organization of Intrinsic Restriction and Modification Genes Bsu M of Bacillus subtilis Marburg

et al. 2002

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“…The E. coli K12 strain C600 F-thi-1 thr-1 pro leuB6 lacY1 tonA21 supE44 hsdR hsdM A- (Maniatis et al 1982) and the B, subtilis strain 1012 hsrM1 leuA8 metB5 (Ikawa et al 1981) were used as host cells for the shuttle vector pHY300PLK (Ishiwa and Shibahara,1985a).…”

Section: Methodsmentioning

confidence: 99%

New shuttle vectors for Escherichia coli and Bacillus subtilis. IV The nucleotide sequence of pHY300PLK and some properties in relation to transformation.

Ishiwa

Shibahara-Sone

1986

The Japanese journal of genetics

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The whole nucleotide sequence of pHY300PLK (Ishiwa and Shibahara 1985a) has been determined, pHY300PLK consists of 4870 by and contains an RNA primer for on-177 and three open reading frames corresponding to TcR, ApR and Rep-al. Twenty-three unique restriction endonuclease recognition sites were found. We have confirmed that ORF al of pHY300PLK is the plasmid replication gene. A rescue phenomenon has been found with respect to multiple infection by pHY300.2PLK (rep-) and pUB110. A series of experiments indicated that the oligomer form, pHY300.2PLK, was rescued by the monomeric form, pUB110. Additionally, some useful information for using pHY300PLK as a shuttle vector in molecular cloning studies is presented.

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“…(hsmF+ hsrF', SP, lysogenic [18]) were grown in TY medium (3). Escherichia coli C600 was grown in L medium (26).…”

Section: Methodsmentioning

confidence: 99%

“…All of the other systems have been introduced in strain 168 through transformation (17,41,44). Genes controlling these systems have been mapped in the B. subtilis 168 chromosome (18). The specificities of the R/M systems BsuR (-HaeIII) and BsuB (-PstI) have been known for some time (4, 41).…”

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confidence: 99%

Restriction and modification in Bacillus subtilis: sequence specificities of restriction/modification systems BsuM, BsuE, and BsuF

Jentsch¹

1983

J Bacteriol

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The sequence specificities of three Bacillus subtilis restriction/modification systems were established: (i) BsuM (CTCGAG), an isoschizomer to XhoI; (ii) BsuE (CGCG), an isoschizomer to FnuDII; and (iii) BsuF (CCGG), an isoschizomer to MspI, HpaII. The BsuM modification enzyme methylates the 3' cytosine of the recognition sequence. The BsuF modification enzyme methylates the 5' cytosine of the sequence, rendering such sites resistant to MspI degradation and leaving the majority of sites sensitive to HpaII degradation.

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Chromosomal loci of genes controlling site-specific restriction endonucleases of Bacillus subtilis

Cited by 24 publications

References 36 publications

Molecular Organization of Intrinsic Restriction and Modification Genes Bsu M of Bacillus subtilis Marburg

Molecular Organization of Intrinsic Restriction and Modification Genes Bsu M of Bacillus subtilis Marburg

New shuttle vectors for Escherichia coli and Bacillus subtilis. IV The nucleotide sequence of pHY300PLK and some properties in relation to transformation.

Restriction and modification in Bacillus subtilis: sequence specificities of restriction/modification systems BsuM, BsuE, and BsuF

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