A manganese-stimulated endonuclease from Bacillus subtilis

Scher, Barbara M.; Dubnau, David

doi:10.1016/0006-291x(73)91185-6

Cited by 21 publications

(18 citation statements)

References 15 publications

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“…2 An additional 60-kDa DNase is an inactive enzyme in the cell-free extract and is encoded in an unidentified gene. 2 Up to date, the presence of several DNases, including a Ca 2ϩ -dependent exonuclease (25), an ATP-dependent nuclease (26,27), and a Mg 2ϩ -dependent endonuclease (28,29), have been reported in B. subtilis. Merchante et al (30) have found several DNases in the periplasm, membrane, and cytoplasm by using zymogram, and Coughlin et al (31) have also analyzed B. subtilis DNases by using two-dimensional zymography analysis and detected 83 nuclease spots.…”

Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a Bacillus subtilis 168 Nuclease, YokF, Involved in Chromosomal DNA Degradation and Cell Death Caused by Thermal Shock Treatments

Sakamoto¹,

Sasaki²,

Tsuchido³

2001

Journal of Biological Chemistry

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We purified and characterized a 39-kDa Bacillus subtilis 168 nuclease that has been suggested in this laboratory to be involved in chromosomal DNA degradation induced by lethal heat and cold shock treatments in vivo. The nuclease activity was inhibited in vitro by aurintricalboxylic acid but not by Zn 2؉ . By the mutant analysis, we identified the 39-kDa nuclease as a product of yokF gene. The yokF gene contained a putative lipoprotein signal peptide motif. After in vivo exposure to lethal heat and cold stresses, the chromosomal DNA fragmentation was reduced in the yokF mutant, which demonstrated about a 2-10-fold higher survival rate than the wild type. The yokF mutant was found to be more sensitive to mitomycin C than the wild type. The transformation efficiency of the yokF mutant was about 10 times higher than that of the wild type. It is suggested that when B. subtilis cells are exposed to a stressful thermal shock resulting in membrane perturbation, YokF nuclease consequently dislocates into the cytoplasm and then attacks DNA.In cells damaged irreversibly by a lethal stress treatment, a variety of structural molecules are subjected to the activated cellular degradation system. This degradation is due to functions of endogenous degradative enzymes, such as autolysins (peptidoglycan hydrolases), proteases, phospholipases, RNases, and DNases.Evidence has shown that a variety of structures and functions in bacterial cell are damaged by heat stress, but the relationship between the damage and cell death is still unclear. Among those, DNA and its functions should be of critical importance for cell survival. Earlier physiological studies have dealt with the effect of heat on the production of single and double strand DNA breaks. In Escherichia coli, the occurrence of single strand breaks at a lethal temperature of 52°C was first demonstrated by Bridges et al. (1), and afterward, it was reported that the double strand break was also incurred on DNA by in vivo heating at the same temperature (2). Although a single strand break has been detected by using the alkaline sucrose gradient technique (3), this technique also picks up apurinic sites in DNA strands (4). In E. coli, endonuclease IV is a representative DNA repair enzyme. After exposure to 52°C, an E. coli mutant defective in this enzyme demonstrates only 20 -30% of the viability of the wild type strain (5). Because endonuclease IV is a major endonuclease acting on apurinic sites in E. coli DNA, it may be involved in the first stage of the DNA excision-repair pathway. The mutant had less DNA breaks after heat treatment, confirming that the production of DNA breaks in this case is part of the DNA repair process.Cold shock has also been reported on E. coli to result in DNA damage as well as cell death (6 -8). It has been suggested that one possible mechanism for cold shock lethality is the loss of magnesium ion from cells, leading to the inactivation of the magnesium-dependent DNA ligase, which joins phosphodeoxyribo-linkage gaps in the strand produced during the...

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Section: Discussionmentioning

confidence: 99%

Purification and Characterization of a Bacillus subtilis 168 Nuclease, YokF, Involved in Chromosomal DNA Degradation and Cell Death Caused by Thermal Shock Treatments

Sakamoto¹,

Sasaki²,

Tsuchido³

2001

Journal of Biological Chemistry

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“…A nuclease apparently similar to the activity described here has been identified and purified from extracts of Bacillus subtilis (Scher & Dubnau, 1973, 1976.…”

Section: Discussionmentioning

confidence: 58%

A Salmonella typhimurium Endonuclease that Converts Native DNA to Fragments of about 8 X 105 daltons

Schumann

Bade

1977

Journal of General Microbiology

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319Crude extracts of Salmonella typhimurium were found to contain an endonuclease that degraded double-stranded linear DNA from bacteria and phages to fragments with a molecular weight of about 8 x 105. The nuclease did not have an absolute requirement for Mg2+. One discrete intermediate product had a molecular weight of 6.6 x I O~. Extracts from two different mutants were tested : one completely lacked the endonuclease activity (strain DB5575), and the other showed an absolute requirement for Mg2+ (strain 4543). No biological role has yet been found for this endonuclease of S. typhimurium.

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“…Although Bacillus subtilis and other bacteria contain an abundance of deoxyribonucleases (Kerr et al, 1965;Birnboim, 1966;McCarthy & Nester, 1969;Ohi & Sueoka, 1973;Scher & Dubnau, 1973 ;Doly & Anagnostopoulos, 1976), their activities are well controlled by protein inhibitors (Strauss & Marone, 1967;McCarthy & Nester, 1969). In non-growing cells and in lysates incubated under physiological conditions, the rate of DNA breakdown is quite low.…”

Section: Introductionmentioning

confidence: 99%

Potentiation of a Nucleolytic Activity in Bacillus subtilis

Sargent¹,

Davies²,

Bennett³

1985

Microbiology

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In several strains of Bacillus subtilis extensive breakdown of chromosomal DNA may be potentiated by osmotic lysis of protoplasts. At its most severe, in strains originating from Farmer & Rothman's thymine auxotroph, the rate of DNA breakdown was greater than 50% per hour at 40 degrees C. The rate of DNA breakdown in most other strains tested was approximately 5% per hour except for SP beta- strains, in which the rate of DNA breakdown was only 0.3%. DNA degradation was attributed to relaxation of control of a nuclease specified by the prophage of SP beta or a related phage. The most potent nuclease in lysates was an ATP-activated protein of Mr 280 000. Derivatives of Farmer and Rothman's strain containing integrated plasmids had the highest rate of DNA degradation. Although the chromosome was completely destroyed, covalently closed circular plasmids were generated from the integrated sequence. These showed massive deletions of the B. subtilis part of the integrated plasmid but the vector sequence remained intact. The nucleolytic activity therefore appears to recognize specific sequences in B. subtilis DNA. We suggest that activation of SP beta genes during development of competence may be a cause of deletion of cloned genes in the early stages of establishment of cloned sequences.

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A manganese-stimulated endonuclease from Bacillus subtilis

Cited by 21 publications

References 15 publications

Purification and Characterization of a Bacillus subtilis 168 Nuclease, YokF, Involved in Chromosomal DNA Degradation and Cell Death Caused by Thermal Shock Treatments

Purification and Characterization of a Bacillus subtilis 168 Nuclease, YokF, Involved in Chromosomal DNA Degradation and Cell Death Caused by Thermal Shock Treatments

A Salmonella typhimurium Endonuclease that Converts Native DNA to Fragments of about 8 X 105 daltons

Potentiation of a Nucleolytic Activity in Bacillus subtilis

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