Cloning and characterization of a DNA region encoding a stress-sensitive restriction system from Corynebacterium glutamicum ATCC 13032 and analysis of its role in intergeneric conjugation with Escherichia coli

Schäfer, Andreas; Schwarzer, A.; Kalinowski, Jörn; Pühler, Alfred

doi:10.1128/jb.176.23.7309-7319.1994

Cited by 54 publications

(33 citation statements)

References 59 publications

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“…CGP3 (185.8 kb, 48.4% GϩC) is adjacent to a cluster of seven tRNA genes. The few recognizable CGP3 functions include the cglIM, cglIR, and cglIIR genes of the C. glutamicum stress-sensitive restrictionmodification system, which is the major barrier to efficient gene transfer into this strain (382,383). The cglIM-specified 5-cytosine methyltransferase apparently methylates the sequence GCSGC (206,382,383).…”

Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

“…The few recognizable CGP3 functions include the cglIM, cglIR, and cglIIR genes of the C. glutamicum stress-sensitive restrictionmodification system, which is the major barrier to efficient gene transfer into this strain (382,383). The cglIM-specified 5-cytosine methyltransferase apparently methylates the sequence GCSGC (206,382,383). Hybridization experiments revealed that the cgl gene region is absent from other C. glutamicum strains and closely related corynebacterial species (383).…”

Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

“…The cglIM-specified 5-cytosine methyltransferase apparently methylates the sequence GCSGC (206,382,383). Hybridization experiments revealed that the cgl gene region is absent from other C. glutamicum strains and closely related corynebacterial species (383). The CGP3 island therefore contributes specifically to the evolution of C. glutamicum ATCC 13032 by coupling the efficiency of HGT to unfavorable environmental conditions.…”

Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

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Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Ventura

Canchaya

Tauch

et al. 2007

Microbiol Mol Biol Rev

946

638

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SUMMARY Actinobacteria constitute one of the largest phyla among Bacteria and represent gram-positive bacteria with a high G+C content in their DNA. This bacterial group includes microorganisms exhibiting a wide spectrum of morphologies, from coccoid to fragmenting hyphal forms, as well as possessing highly variable physiological and metabolic properties. Furthermore, Actinobacteria members have adopted different lifestyles, and can be pathogens (e.g., Corynebacterium, Mycobacterium, Nocardia, Tropheryma, and Propionibacterium), soil inhabitants (Streptomyces), plant commensals (Leifsonia), or gastrointestinal commensals (Bifidobacterium). The divergence of Actinobacteria from other bacteria is ancient, making it impossible to identify the phylogenetically closest bacterial group to Actinobacteria. Genome sequence analysis has revolutionized every aspect of bacterial biology by enhancing the understanding of the genetics, physiology, and evolutionary development of bacteria. Various actinobacterial genomes have been sequenced, revealing a wide genomic heterogeneity probably as a reflection of their biodiversity. This review provides an account of the recent explosion of actinobacterial genomics data and an attempt to place this in a biological and evolutionary context.

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Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

Section: Prophage-like Elements In the C Glutamicum Genomementioning

confidence: 99%

See 1 more Smart Citation

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Ventura

Canchaya

Tauch

et al. 2007

Microbiol Mol Biol Rev

946

638

View full text Add to dashboard Cite

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“…Accordingly, there are many examples in which conjugal transfer is unaffected by restriction barriers that prevent the infection of unmodified phage or the transformation of unmodified DNA (5,20,36,41,55,66). However, there are also reports of restriction in nonenteric bacteria indeed limiting conjugal efficiency (18,44,64,79).Regardless of the biological importance of restriction, the resulting inefficiency of DNA transfer is a frequently encountered barrier against introducing DNA into bacteria of experimental interest. While restriction may reduce the frequency of DNA transfer below levels of detectability (40,42,79), it is also possible that some introduced DNA with unmodified restriction sites may escape destruction.…”

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Reduction of conjugal transfer efficiency by three restriction activities of Anabaena sp. strain PCC 7120

et al. 1997

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The efficiency of conjugal transfer of plasmids from Escherichia coli to the cyanobacterium Anabaena sp. strain PCC 7120 was quantitated as a function of the number of restriction sites for the restriction enzymes carried by the recipient. In addition to the previously recognized isoschizomers of AvaI and AvaII, PCC 7120 was found to possess an isoschizomer of AvaIII. Plasmids modified in E. coli with methylases that protect in vitro against restriction by the three enzymes were transferred with high efficiency, nearly independent of the number of restriction sites on the plasmid. Plasmids left unprotected against one of the three restriction enzymes were transferred with lower efficiencies. For low numbers of sites, the efficiency of conjugal transfer decreased as an exponential function of the number of unprotected sites. The methods presented may be used to increase the efficiency of conjugal transfer into restriction-competent bacteria.Since first recognized, restriction in vivo has been most closely identified with the reduction of plating efficiency of bacteriophage (7, 10). Although conjugal transfer of unmodified DNA also was shown to be sensitive to restriction in Escherichia coli (10), the inability of certain restriction enzymes to act on single-stranded DNA (49, 56) has led to the view that conjugally transferred single-stranded DNA (37) should be immune from restriction (5, 69). Accordingly, there are many examples in which conjugal transfer is unaffected by restriction barriers that prevent the infection of unmodified phage or the transformation of unmodified DNA (5,20,36,41,55,66). However, there are also reports of restriction in nonenteric bacteria indeed limiting conjugal efficiency (18,44,64,79).Regardless of the biological importance of restriction, the resulting inefficiency of DNA transfer is a frequently encountered barrier against introducing DNA into bacteria of experimental interest. While restriction may reduce the frequency of DNA transfer below levels of detectability (40,42,79), it is also possible that some introduced DNA with unmodified restriction sites may escape destruction. With this in mind, it is often of practical importance to know quantitatively the degree to which each restriction site impairs DNA transfer into restriction-competent recipients.The effect of restriction sites on the efficiency of DNA transfer has been quantitated in several studies using plasmids or phage with a known number of sites. Such studies have measured the effect of restriction on phage infection (47, 48, 54), phage transfection (35), transformation (17,51,71), and electroporation (8,34,43). Reduction of conjugal transfer efficiency by restriction has not been systematically studied, although there have been some quantitative reports (30,70 (14,18,42). We report here the construction of plasmids carrying methylases that protect against one or more restriction activities of Anabaena sp. strain PCC 7120, including a third restriction activity reported here for the first time. Using these plasmids, ...

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“…SOS induction, in turn, reduces by 1 or more orders of magnitude the activities of type I enzymes and of the methylation-dependent restriction endonuclease McrBC (20,23,53). Likewise, a restriction system encoded by plasmid RP4 is inactivated by various environmental stress factors (44).…”

Section: Figmentioning

confidence: 99%

Short-range and long-range context effects on coliphage T4 endonuclease II-dependent restriction

1996

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Synthetic sites inserted into a plasmid were used to analyze the sequence requirements for in vivo DNA cleavage dependent on bacteriophage T4 endonuclease II. A 16-bp variable sequence surrounding the cleavage site was sufficient for cleavage, although context both within and around this sequence influenced cleavage efficiency. The most efficiently cleaved sites matched the sequence CGRCCGCNTTGGCNGC, in which the strongly conserved bases to the left were essential for cleavage. The less-conserved bases in the center and in the right half determined cleavage efficiency in a manner not directly correlated with the apparent base preference at each position; a sequence carrying, in each of the 16 positions, the base most preferred in natural sites in pBR322 was cleaved infrequently. This, along with the effects of substitutions at one or two of the lessconserved positions, suggests that several combinations of bases can fulfill the requirements for recognition of the right part of this sequence. The replacements that improve cleavage frequency are predicted to influence helical twist and roll, suggesting that recognition of sequence-dependent DNA structure and recognition of specific bases are both important. Upon introduction of a synthetic site, cleavage at natural sites within 800 to 1,500 bp from the synthetic site was significantly reduced. This suggests that the enzyme may engage more DNA than its cleavage site and cleaves the best site within this region. Cleavage frequency at sites which do not conform closely to the consensus is, therefore, highly context dependent. Models and possible biological implications of these findings are discussed.Sequence-specific endonucleases are crucial for the proper maintenance of DNA in cells; they are required for its replication, recombination, and repair; its defense against invading parasites; and its proper degradation during programmed cell death in differentiated organisms. Scrutiny reveals strong similarities in their mechanisms of action, suggesting that some may participate in more than one such event.The restriction system encoded by coliphage T4 and dependent on its endonuclease II (EndoII) resembles type II restriction endonucleases more than type I or III or the methylationdependent endonucleases. Restriction enzymes recognize their specific DNA targets by a combination of direct and indirect readout (30, 31), i.e., interactions between nucleotide bases and amino acid residues and interactions between amino acids and the sugar-phosphate backbone, respectively. Both interactions are dependent on the sequence of the recognition site. Our previous investigations of cleavage in vivo of unmodified T4 DNA (24) as well as of a small plasmid (pBR322 [7]) suggested that recognition and cleavage by EndoII were dependent on helical structure. The helical structure of the recognition site may be influenced by factors outside the consensus sequence and may, in turn, influence the tracking of a processive enzyme along the DNA molecule.However, several features of T4 EndoII ...

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Cloning and characterization of a DNA region encoding a stress-sensitive restriction system from Corynebacterium glutamicum ATCC 13032 and analysis of its role in intergeneric conjugation with Escherichia coli

Cited by 54 publications

References 59 publications

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Genomics ofActinobacteria: Tracing the Evolutionary History of an Ancient Phylum

Reduction of conjugal transfer efficiency by three restriction activities of Anabaena sp. strain PCC 7120

Short-range and long-range context effects on coliphage T4 endonuclease II-dependent restriction

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