Date of Award

Summer 1981

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

Program/Concentration

Chemistry

Committee Director

Thomas O. Sitz

Committee Member

James H. Yuan

Committee Member

Ken Somers

Call Number for Print

Special Collections LD4331.C45 G34

Abstract

An assay procedure for in vitro enzymatic methylation of mannnalian ribosomal RNA has been developed in this study. The assay procedure, utilized for the comparison of normal and neoplastic methylase activities (using mouse liver and Ehrlich ascites cells as sources of enzyme), is a modification of previously published methods (52,53). A 100,000 x g supernatant (SlOO) enzyme preparation was incubated with 28S-5.8S rRNA and tritium-labeled S-adenosyl-L-methionine. The RNA was extracted, applied to DEAE cellulose paper, washed, and the radioactivity counted. The neoplastic cell methylase preparation was more active in methylating both exogenous neoplastic and normal 28S-5.8S rRNA than the normal enzyme preparation. However, based on DEAESephadex chromatograms of ribonuclease T2 digests of tritium-labeled RNA from neoplastic cell methylase assays, the in vitro methylation is almost exclusively restricted to the purine and pyrimidine bases. Although this in vitro base methylation may not represent the cellular situation, which is almost exclusively 2'-O-ribose methylation, the higher neoplastic cell methylase activity correlates well with previously published studies and indicates that this assay procedure is a useful tool for further study of normal and neoplastic cell methylase activities.

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DOI

10.25777/2k2f-yh44

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