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. 1969 Mar;97(3):1426–1430. doi: 10.1128/jb.97.3.1426-1430.1969

Heat-activated Endonuclease in Bacillus subtilis

Charlotte McCarthy 1,2,1, Eugene W Nester 1,2
PMCID: PMC249864  PMID: 4975749

Abstract

A heat-stable, heat-activated endonuclease was found in sonic extracts of the transformable strain Bacillus subtilis 168 when the organism was grown to logarithmic phase in minimal medium. The enzyme was not present in the poorly transformable strain 23. The endonuclease was stable to 100 C for 30 min and, in a crude extract, was activated by heating at 80, 90, or 100 C. The activation caused a 5- to 10-fold increase in total units of enzyme activity. Sucrose gradient centrifugation indicated that the enzyme in a crude preparation has a major form (molecular weight, 66,000) which remains unchanged after heat activation. Under the assay conditions employed, the endonuclease did not release acid-soluble material from the substrate, high molecular weight tritiated deoxyribonucleic acid. The product, in double-stranded form, had a molecular weight of approximately 105, but it appeared to have undergone single-strand breaks.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BURGI E., HERSHEY A. D. Sedimentation rate as a measure of molecular weight of DNA. Biophys J. 1963 Jul;3:309–321. doi: 10.1016/s0006-3495(63)86823-x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Birnboim H. C. Cellular site in Bacillus subtilis of a nuclease which preferentially degrades single-stranded nucleic acids. J Bacteriol. 1966 Mar;91(3):1004–1011. doi: 10.1128/jb.91.3.1004-1011.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duncan J. L., Groman N. B. Studies of the activity of diphtheria toxin. I. Poliovirus replication in intoxicated HeLa cells. J Exp Med. 1967 Mar 1;125(3):489–500. doi: 10.1084/jem.125.3.489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Farmer J. L. Deoxyribonucleic acid degradation in Bacillus subtilis during exposure to actinomycin D. J Bacteriol. 1968 Apr;95(4):1450–1460. doi: 10.1128/jb.95.4.1450-1460.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. GEIDUSCHEK E. P., DANIELS A. A SIMPLE ASSAY FOR DNA ENDONUCLEASES. Anal Biochem. 1965 Apr;11:133–137. doi: 10.1016/0003-2697(65)90052-7. [DOI] [PubMed] [Google Scholar]
  7. Kerr I. M., Pratt E. A., Lehman I. R. Exonucleolytic degradation of high-molecular-weight DNA and RNA to nucleoside 3'-phosphates by a nuclease from B. subtilis. Biochem Biophys Res Commun. 1965 Jul 12;20(2):154–162. doi: 10.1016/0006-291x(65)90339-6. [DOI] [PubMed] [Google Scholar]
  8. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  9. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  10. NAKAI M., MINAMI Z., YAMAZAKI T., TSUGITA A. STUDIES ON THE NUCLEASES OF A STRAIN OF BACILLUS SUBTILIS. J Biochem. 1965 Jan;57:96–99. doi: 10.1093/oxfordjournals.jbchem.a128063. [DOI] [PubMed] [Google Scholar]
  11. Okazaki R., Okazaki T., Sakabe K. An extracellular nuclease of Bacillus subtilis: some novel properties as a DNA exonuclease. Biochem Biophys Res Commun. 1966 Mar 22;22(6):611–619. doi: 10.1016/0006-291x(66)90190-2. [DOI] [PubMed] [Google Scholar]
  12. SMEATON J. R., ELLIOTT W. H., COLEMAN G. AN INHIBITOR IN BACILLUS SUBTILIS OF ITS EXTRACELLULAR RIBONUCLEASE. Biochem Biophys Res Commun. 1965 Jan 4;18:36–42. doi: 10.1016/0006-291x(65)90878-8. [DOI] [PubMed] [Google Scholar]
  13. Spizizen J. TRANSFORMATION OF BIOCHEMICALLY DEFICIENT STRAINS OF BACILLUS SUBTILIS BY DEOXYRIBONUCLEATE. Proc Natl Acad Sci U S A. 1958 Oct 15;44(10):1072–1078. doi: 10.1073/pnas.44.10.1072. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Strauss B., Marone R. A heat-labile inhibitor of deoxyribonucleic acid degradation in Bacillus subtilis. Biochem Biophys Res Commun. 1967 Oct 11;29(1):143–147. doi: 10.1016/0006-291x(67)90555-4. [DOI] [PubMed] [Google Scholar]

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