A new pyrimidine-specific ribonuclease from bovine seminal plasma which is active on both single and double-stranded polyribonucleotides and which can distinguish between Mg2+-containing and Mg2+ -depleted naturally occurring RNAs

E S P REDDY1, K H SCHEIT2, N SITARAM1 & P M BHARGAVA1

1Centre for Cellular and Molecular Biology Regional Research Laboratory Hyderabad 500009, India.

2Max-Planck Institut für Biophysikalische Chemie Abteilung Moleukulare Biologie Am Fassberg, D-3400 Goettingen-Nikolausberg Federal Republic of Germany

Published in: Journal of Molecular Biology, 1979, 135, 525-544.

Abstract:

The purification to homogeneity of a new ribonuclease, named RNAase SPL, from bovine seminal plasma is described. This nuclease, like the bovine pancreatic RNAase A, is pyrimidine specific. Its activity on single-stranded synthetic polyribonucleotides such as poly(rU) is significantJy higher than that of RNAase A. However, unlike RNAase A, RNAase SPL is highly active on a double-stranded RNA such as poly[r(A . U)], and shows extremely limited activity on naturally occurring RNAs, such as Escherichia coli RNA, prepared with Mg2+ present throughout the isolation procedure. Under conditions of limiting hydrolysis in which RNAase A degrades 60 to 90% of total E. coli RNA to acid-soluble material and the remaining to material having a molecular weight lower than that of transfer RNA, RNAase SPL does not yield any acid-soluble products: it does not appear to degrade tRNA or 5 S RNA, and causes only a small number of nicks in the remaining RNAs to yield a limiting digest containing products with molecular weights ranging between 10,000 and 150,000. Absence of Mg2+ during the isolation procedure, or heat denaturation of the RNA makes it as susceptible to RNAase SPL as it is to RNAase A.

The above and other related observations reported here support the view that there are Mg2+ -dependent structural features, besides single and double-strandedness, in naturally occurring RNAs, that can be distinguished by using the two nucleases RNAase SPL and RNAase A.

PMID: 537087 | DOI: 10.1016/0022-2836(79)90162-1

Keywords:
RNAase A, bovine pancreatic ribonuclease; RNAase SPL, seminal plasma ribonuclease

HOW TO CITE

A new pyrimidine-specific ribonuclease from bovine seminal plasma which is active on both single and double-stranded polyribonucleotides and which can distinguish between Mg2+-containing and Mg2+ -depleted naturally occurring RNAs. E S P REDDY, K H SCHEIT, N SITARAM & P M BHARGAVA. Journal of Molecular Biology, 1979, 135, 525-544.

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