PMID:1704882
| Citation |
Padmanabha, KP and Deutscher, MP (1991) RNase T affects Escherichia coli growth and recovery from metabolic stress. J. Bacteriol. 173:1376-81 |
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| Abstract |
To determine the essentiality and role of RNase T in RNA metabolism, we constructed an Escherichia coli chromosomal rnt::kan mutation by using gene replacement with a disrupted, plasmid-borne copy of the rnt gene. Cell extracts of a strain with mutations in RNases BN, D, II, and I and an interuppted rnt gene were devoid of RNase T activity, although they retained a low level (less than 10%) of exonucleolytic activity on tRNA-C-C-[14C]A due to two other unidentified RNases. A mutant lacking tRNA nucleotidyltransferase in addition to the aforementioned RNases accumulated only about 5% as much defective tRNA as did RNase T-positive cells, indicating that this RNase is responsible for essentially all tRNA end turnover in E. coli. tRNA from rnt::kan strains displayed a slightly reduced capacity to be aminoacylated, raising the possibility that RNase T may also participate in tRNA processing. Strains devoid of RNase T displayed slower growth rates than did the wild type, and this phenotype was accentuated by the absence of the other exoribonucleases. A strain lacking RNase T and other RNases displayed a normal response to UV irradiation and to the growth of bacteriophages but was severely affected in its ability to recover from a starvation regimen. The data demonstrate that the absence of RNase T affects the normal functioning of E. coli, but it can be compensated for to some degree by the presence of other RNases. Possible roles of RNase T in RNA metabolism are discussed. |
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| Keywords |
Blotting, Southern; Cell Division/physiology; Chromatography, Gel; Cloning, Molecular; DNA/analysis; Escherichia coli/genetics; Escherichia coli/physiology; Exoribonucleases/physiology; Gene Expression; Plasmids; RNA/metabolism; Restriction Mapping; Transformation, Genetic |
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