Abstract
A nuclease from N. crassa has been prepared to the hydroxylapatite stage of purification described by Rabin and Fraser (1). It degrades single stranded DNA in an essentially exonucleolytic process. It does not give any appreciable acid soluble material with double stranded DNA as substrate. This shows its high degree of specificity towards single stranded DNA.
An extra activity which makes double strand breaks in the AT rich regions of DNA is described. A limited number of breaks are observed with calf or mouse DNAs, as judged by the decrease of sedimentation velocity in alkaline or neutral sucrose gradients. The number of breaks decreases with increasing the ionic strength. λ DNA is broken in half at the AT rich middle region of the molecule. The circular replicative form of fd DNA is converted to linear pieces of a rather homogeneous length. It is concluded from these results that it is an endonuclease activity specific for d(A-T) rich regions in double stranded DNA.
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Selected References
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- Aaij C., Borst P. The gel electrophoresis of DNA. Biochim Biophys Acta. 1972 May 10;269(2):192–200. doi: 10.1016/0005-2787(72)90426-1. [DOI] [PubMed] [Google Scholar]
- Ando T. A nuclease specific for heat-denatured DNA in isolated from a product of Aspergillus oryzae. Biochim Biophys Acta. 1966 Jan 18;114(1):158–168. doi: 10.1016/0005-2787(66)90263-2. [DOI] [PubMed] [Google Scholar]
- Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
- Godson G. N. Action of the single-stranded DNA specific nuclease S1 on double-stranded DNA. Biochim Biophys Acta. 1973 Apr 21;308(7):59–67. doi: 10.1016/0005-2787(73)90122-6. [DOI] [PubMed] [Google Scholar]
- HERSHEY A. D., BURGI E. COMPLEMENTARY STRUCTURE OF INTERACTING SITES AT THE ENDS OF LAMBDA DNA MOLECULES. Proc Natl Acad Sci U S A. 1965 Feb;53:325–328. doi: 10.1073/pnas.53.2.325. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Johnson P. H., Laskowski M., Sr Mung bean nuclease I. II. Resistance of double stranded deoxyribonucleic acid and susceptibility of regions rich in adenosine and thymidine to enzymatic hydrolysis. J Biol Chem. 1970 Feb 25;245(4):891–898. [PubMed] [Google Scholar]
- LINN S., LEHMAN I. R. AN ENDONUCLEASE FROM NEUROSPORA CRASSA SPECIFIC FOR POLYNUCLEOTIDES LACKING AN ORDERED STRUCTURE. II. STUDIES OF ENZYME SPECIFICITY. J Biol Chem. 1965 Mar;240:1294–1304. [PubMed] [Google Scholar]
- Rabin E. Z., Fraser M. J. Isolation of Neurospora crassa endonuclease specific for single-stranded DNA. Can J Biochem. 1970 Mar;48(3):389–392. doi: 10.1139/o70-063. [DOI] [PubMed] [Google Scholar]
- Rabin E. Z., Preiss B., Fraser M. J. A nuclease from Neurospora crassa conidia specific for single-stranded nucleic acids. Prep Biochem. 1971;1(4):283–307. doi: 10.1080/00327487108081946. [DOI] [PubMed] [Google Scholar]
- Rabin E. Z., Tenenhouse H., Fraser M. J. An exonuclease of Neurospora crassa specific for single-stranded nucleic acids. Biochim Biophys Acta. 1972 Jan 18;259(1):50–68. doi: 10.1016/0005-2787(72)90473-x. [DOI] [PubMed] [Google Scholar]
- STUDIER F. W. SEDIMENTATION STUDIES OF THE SIZE AND SHAPE OF DNA. J Mol Biol. 1965 Feb;11:373–390. doi: 10.1016/s0022-2836(65)80064-x. [DOI] [PubMed] [Google Scholar]
- Skalka A., Burgi E., Hershey A. D. Segmental distribution of nucleotides in the DNA of bacteriophage lambda. J Mol Biol. 1968 May 28;34(1):1–16. doi: 10.1016/0022-2836(68)90230-1. [DOI] [PubMed] [Google Scholar]
- Sutton W. D. A crude nuclease preparation suitable for use in DNA reassociation experiments. Biochim Biophys Acta. 1971 Jul 29;240(4):522–531. doi: 10.1016/0005-2787(71)90709-x. [DOI] [PubMed] [Google Scholar]
- Wingert L., Von Hippel P. H. The conformation dependent hydrolysis of DNA by micrococcal nuclease. Biochim Biophys Acta. 1968 Mar 18;157(1):114–126. doi: 10.1016/0005-2787(68)90270-0. [DOI] [PubMed] [Google Scholar]