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Gene affecting longevity of messenger RNA: a mutant of Escherichia coli with altered mRNA stability.
Authors:Michihiko Kuwano   Mayumi Ono   Hideya Endo   Katsuji Hori   Kunihiko Nakamura   Yukinori Hirota  Yoshinari Ohnishi
Affiliation:(1) Cancer Research Institute, Faculty of Medicine, Kyushu University, 812 Fukuoka;(2) Division of Molecular Genetics, Department of Biology, Faculty of Science, Kyushu University, 812 Fukuoka;(3) Department of Bacteriology, Faculty of Medicine, Kagoshima University, 892 Kagoshima;(4) National Institute of Genetics, 411 Mishima;(5) Department of Bacteriology, School of Medicine, Tokushima University, T70 Tokushima, Japan
Abstract:Summary we have screened 897 temperature sensitive growth mutants ofE. coli for mutant strains showing longer mRNA half-life. The fate of pulse-labelled RNA was examined at 42° C after cessation of RNA synthesis and with prior exposure to nonpermissive temperature (42° C). Eight stains showed altered turn-over of RNA (presumably mRNA), and further analysis on mutant strain JE15144 indicated that the stability of pulse-labeled RNA as well as of tryptophan (trp) mRNA increased four to seven fold over its parental strain at 42° C. At 4 min or 10 min after addition of rifampicin, some 70 to 80% of polyribosome in the growing cells could still be conserved in JE15144 cultured at the nonpermissive temperature while little, if any, polyribosomes remained in its parental strain (PA3092) under the same condition. Two generation times were required for complete stoppage of growth of this mutant strain after shifting to 42° C, and protein synthesis continued at a significant, but slightly reduced, rate at 42° C. However, functional decay of mRNA in the mutant strain, with respect to the capacity for producing peptides, appeared to be similar to the parent strain, with half-lives of 3.5 min in PA3092 and 4.7 min in JE15144.
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