New Sequence-specific Human Ribonuclease: Purification and Properties

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 1998 Aug 15

PMID 9705518

Citations 2

Authors

G Przewlocki

J Lipecka

A Edelman

A Przykorska

Affiliations

Soon will be listed here.

Abstract

A new sequence-specific RNase was isolated from human colon carcinoma T84 cells. The enzyme was purified to electrophoretical homogeneity by pH precipitation, HiTrapSP and Superdex 200 FPLC. The molecular weight of the new enzyme, which we have named RNase T84, is 19 kDa. RNase T84 is an endonuclease which generates 5'-phosphate-terminated products. The new RNase selectively cleaved the phosphodiester bonds at AU or GU steps at the 3' side of A or G and the 5' side of U. 5'AU3' or 5'GU3' is the minimal sequence required for T84 RNase activity, but the rate of cleavage depends on the sequence and/or structure context. Synthetic ribohomopolymers such as poly(A), poly(G), poly(U) and poly(C) were very poorly hydrolysed by T84 enzyme. In contrast, poly(I) and heteroribopolymers poly(A,U) and poly(A,G,U) were good substrates for the new RNase. The activity towards poly(I) was stronger in two colon carcinoma cell lines than in three other epithelial cell lines. Our results show that RNase T84 is a new sequence-specific enzyme whose gene is abundantly expressed in human colon carcinoma cell lines.

Citing Articles

Molecular cloning and characterization of the human RNase kappa, an ortholog of Cc RNase.

Economopoulou M, Fragoulis E, Sideris D Nucleic Acids Res. 2007; 35(19):6389-98.

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Probing the substrate specificity of Escherichia coli RNase E using a novel oligonucleotide-based assay.

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