Purification and characterization of a major endonuclease from rat liver nuclei

Gerald A. Cordis, Peter J. Goldblatt, Murray P Deutscher

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

A major endonuclease has been purified approximately 800-fold from rat liver nuclei using poly(A) as substrate. The enzyme had a molecular weight of about 50,000, and active fractions were obtained which contained no nucleic acid. Enzymatic activity was optimal between pH 6 and 7 and was totally dependent on the presence of a divalent cation. The reaction was inhibited by high ionic strength, polydextran sulfate, heparin, and sodium pyrophosphate. The purified enzyme readily hydrolyzed poly(A), poly(U), poly(C), and denatured DNA, whereas poly(G) was not degraded, and transfer RNA, ribosomal RNA, and native DNA were hydrolyzed only at relatively slow rates. These data suggest that the enzyme may be specific for single-stranded polynucleotides. The purified enzyme was essentially devoid of exonuclease activity, and the products of exhaustive endonuclease digestion of poly(A) were small oligonucleotides terminated with a 5′-phosphoryl group. Evidence was obtained that this endonuclease is localized in the nucleoplasm. Possible functions for this activity are discussed.

Original languageEnglish
Pages (from-to)2596-2603
Number of pages8
JournalBiochemistry
Volume14
Issue number12
StatePublished - Dec 1 1975
Externally publishedYes

Fingerprint

Endonucleases
Liver
Purification
Rats
Poly A
Enzymes
Poly A-U
Poly G
Poly C
Poly U
Polynucleotides
Exonucleases
Ribosomal RNA
DNA
Divalent Cations
Transfer RNA
Ionic strength
Oligonucleotides
Osmolar Concentration
Nucleic Acids

ASJC Scopus subject areas

  • Biochemistry

Cite this

Cordis, G. A., Goldblatt, P. J., & Deutscher, M. P. (1975). Purification and characterization of a major endonuclease from rat liver nuclei. Biochemistry, 14(12), 2596-2603.

Purification and characterization of a major endonuclease from rat liver nuclei. / Cordis, Gerald A.; Goldblatt, Peter J.; Deutscher, Murray P.

In: Biochemistry, Vol. 14, No. 12, 01.12.1975, p. 2596-2603.

Research output: Contribution to journalArticle

Cordis, GA, Goldblatt, PJ & Deutscher, MP 1975, 'Purification and characterization of a major endonuclease from rat liver nuclei', Biochemistry, vol. 14, no. 12, pp. 2596-2603.
Cordis GA, Goldblatt PJ, Deutscher MP. Purification and characterization of a major endonuclease from rat liver nuclei. Biochemistry. 1975 Dec 1;14(12):2596-2603.
Cordis, Gerald A. ; Goldblatt, Peter J. ; Deutscher, Murray P. / Purification and characterization of a major endonuclease from rat liver nuclei. In: Biochemistry. 1975 ; Vol. 14, No. 12. pp. 2596-2603.
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