A Nuclease from Neurospora Crassa Conidia Specific for Single-Stranded Nucleic Acids
- 3 January 1971
- journal article
- research article
- Published by Taylor & Francis in Preparative Biochemistry
- Vol. 1 (1) , 283-307
- https://doi.org/10.1080/00327487108081946
Abstract
This report describes the purification from sonicates of Neurospora crassa conidia of a nuclease with extremely high specificity for single-stranded nucleic acids. The enzyme was purified 510-fold from streptomycin-treated sonicates in successive steps by (NH4)2SO4 fractionation, acetone fractionation, by chromatography on phosphocellulose, DEAE-cellulose, Sephadex G-200 and hydroxy apatite and, finally, by preparative polyacrylamide gel electrophoresis. The yield of purified enzyme was 7%. Only one protein component was detected by analytical polyacrylamide gel electrophoresis at pH8.9, but, in the presence of 1% sodium dodecyl sulfate and 1% mercaptoethanol at pH7.0, one minor component (approximately 10% of the total protein, mol. wt. approximately 77,000) and one major component (mol. wt. approximately 72,000) were detected. The enzyme degraded denatured DNA rapidly but did not release any acid-soluble material from native DNA. It also did not alter the sedimentation properties of native bacteriophage T7 DNA. The only action on native DNA that was detected was a slow conversion of the superhelical form of bacteriophage S13 DNA to the open circle form. The products of a 10% digest (10% acid-soluble material) of denatured DNA contained 5′-mono-nucleotides and oligonucleotides (di- to decanucleotides) in a ratio of 3 to 1, indicating that the digestion was predominantly exonucleolytic in character.Keywords
This publication has 22 references indexed in Scilit:
- The Synthesis and Breakdown of Nucleic Acids in Mammalian Cells Transformed by Oncogenic VirusesPublished by Elsevier ,1970
- Isolation of Escherichia coli transfer RNA-gene hybridsJournal of Molecular Biology, 1970
- Mung Bean Nuclease IPublished by Elsevier ,1970
- Isolation of Pure lac Operon DNA*Nature, 1969
- Enzymatic Breakage of Deoxyribonucleic AcidPublished by Elsevier ,1969
- A GENETIC ASSAY FOR MRNA'S OF PHAGE T4Proceedings of the National Academy of Sciences, 1969
- Sheep Kidney NucleaseEuropean Journal of Biochemistry, 1967
- An Endonuclease from Neurospora crassa Specific for Polynucleotides Lacking an Ordered StructureJournal of Biological Chemistry, 1965
- An Endonuclease from Neurospora crassa Specific for Polynucleotides Lacking an Ordered StructurePublished by Elsevier ,1965
- Characterization of a deoxyribonuclease of Mustelus canis liverBiochimica et Biophysica Acta (BBA) - Enzymology and Biological Oxidation, 1965