An rne-1 pnp-7 double mutation suppresses the temperature-sensitive defect of lacZ gene expression in a divE mutant.
A divE mutant, which has a temperature-sensitive mutation in the tRNA1Ser gene, exhibits differential loss of the synthesis of certain proteins, such as beta-galactosidase and succinate dehydrogenase, at nonpermissive temperatures. In Escherichia coli, the UCA codon is recognized only by tRNA1Ser. Several genes containing UCA codons are normally expressed after a temperature shift to 42 degrees C in the divE mutant. Therefore, it is unlikely that the defect in protein synthesis at 42 degrees C is simply caused by a defect in the decoding function of the mutant tRNA1Ser. In this study, we sought to determine the cause of the defect in lacZ gene expression in the divE mutant. It has also been shown that the defect in lacZ gene expression is accompanied by a decrease in the amount of lacZ mRNA. To examine whether inactivation of mRNA degradation pathways restores the defect in lacZ gene expression, we constructed divE mutants containing rne-1, rnb-500, and pnp-7 mutations in various combinations. We found that the defect was almost completely restored by introducing an rne-1 pnp-7 double mutation into the divE mutant. Northern hybridization analysis showed that the rne-1 mutation stabilized lacZ mRNA, whereas the pnp-7 mutation stabilized mutant tRNA1Ser, at 44 degrees C. We present a mechanism that may explain these results.
Aiso
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Department of Molecular Biology, School of Health Sciences, Kyorin University, Hachioji, Tokyo, Japan.
Ohki
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R
D000483:Alleles; D015152:Blotting, Northern; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D004251:DNA Transposable Elements; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D015870:Gene Expression; D007763:Lac Operon; D016254:Mutagenesis, Insertional; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D012362:RNA, Transfer, Ser / Q000235:genetics* / Q000378:metabolism; D012260:Ribonucleases / Q000235:genetics; D013385:Succinate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D003062:Codon; D004251:DNA Transposable Elements; D012333:RNA, Messenger; D012362:RNA, Transfer, Ser; D013385:Succinate Dehydrogenase; D012260:Ribonucleases; D001616:beta-Galactosidase