Degradation of RNA in Nuclei From Ehrlich Ascites Cells

Author:

Speers E. Ann,Tigerstrom Richard G. von

Abstract

Nuclei were prepared from Ehrlich ascites cells in 80% yield by homogenization of the cells in an aqueous solution containing Triton N-101 and washing of the nuclear fraction by centrifugation and resuspension. Compared to the enzyme activities present in cell extracts, approximately 47% exo-RNase I, 15% alkaline RNase II, 9% acid RNase II and 7% acid phosphatase were associated with the nuclear fraction after isolation. Exo-RNase I and alkaline RNase II were rapidly lost from nuclei during incubation at 37 °C. The degradation of newly synthesized RNA in nuclei incubated at 37 °C was followed by polyacrylamide gel electrophoresis and by characterization of acid-soluble degradation products. The rate of hydrolysis of the nuclear RNA was rapid during the initial stages of incubation and then proceeded at a much reduced rate. Nucleoside 5′-phosphates were the major acid-soluble degradation products, in agreement with the presence of exo-RNase I. Although a considerable amount of alkaline RNase II was associated with the nuclear fraction, extensive endonucleolytic cleavage of the nuclear RNA was not apparent. Compared to the processing of nuclear RNA in whole cells, however, the degradation in isolated nuclei was relatively non-specific.

Publisher

Canadian Science Publishing

Subject

General Medicine

Cited by 5 articles. 订阅此论文施引文献 订阅此论文施引文献,注册后可以免费订阅5篇论文的施引文献,订阅后可以查看论文全部施引文献

1. On the Mechanism of Reutilization of Nuclear RNA Degradation Products: A Study in Vivo;Metabolism and Enzymology of Nucleic Acids;1988

2. BIBLIOGRAPHY;Kinetics of Experimental Tumour Processes;1982

3. Electrophoretic investigation of nuclear membrane ribonucleases of rat liver and hepatoma-27 cells;Bulletin of Experimental Biology and Medicine;1977-12

4. Mammalian Nucleolytic Enzymes;Progress in Nucleic Acid Research and Molecular Biology;1977

5. Isolation and characterization of two alkaline ribonucleases from calf serum;Biochimica et Biophysica Acta (BBA) - Nucleic Acids and Protein Synthesis;1976-01

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