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. 1982 Sep;151(3):1162–1170. doi: 10.1128/jb.151.3.1162-1170.1982

Characterization of polyadenylated RNA in a protein-producing bacterium, Bacillus brevis 47.

I Hussain, N Tsukagoshi, S Udaka
PMCID: PMC220392  PMID: 6179922

Abstract

Up to about 50% of the total radioactivity in pulse-labeled RNA in Bacillus brevis 47-5, a high-protein-producing bacterium, was found in the polyadenylated fraction [termed poly(A)-RNA] isolated by adsorption to oligodeoxythymidylic acid-cellulose. Labeled RNA was bound to the cellulose regardless of whether the radioactive precursor was [3H]adenosine or [3H]uridine, showing that the adsorbed material was poly(A)-RNA rather than free poly(A). Poly(A) tracts, isolated after digestion of pulse-labeled RNA with pancreatic and T1 RNases, were homogeneous, with a length of about 95 nucleotides. Susceptibility of the isolated poly(A) tracts to degradation by snake venom phosphodiesterase and polynucleotide phosphorylase indicated that the poly(A) sequences were located directly at the 3'-terminal of the RNA molecules. Comparison of the poly(A)-RNA content in high-protein-producing and nonprotein-producing cells of B. brevis 47 showed much higher levels in the former. Electrophoretic analysis in both denaturing and denaturing polyacrylamide gels of the poly(A)-RNAs showed a heterogeneous population of molecules ranging in size from 23S to 4S. Comparison of the molecular-weight distribution patterns revealed that a significantly greater amount of high-molecular-weight poly(A)-RNA (comigrating with 23S RNA) was present under conditions in which extracellular protein production was high. The possibility that a substantial fraction of the poly(A)-RNA might be involved in the synthesis of extracellular proteins in B. brevis 47 is discussed.

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Selected References

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