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  • American Society for Microbiology  (3)
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  • American Society for Microbiology  (3)
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  • 1
    Online Resource
    Online Resource
    American Society for Microbiology ; 2013
    In:  Journal of Bacteriology Vol. 195, No. 9 ( 2013-05), p. 2039-2049
    In: Journal of Bacteriology, American Society for Microbiology, Vol. 195, No. 9 ( 2013-05), p. 2039-2049
    Abstract: We have performed a screening of hydroxyurea (HU)-sensitive mutants using a single-gene-deletion mutant collection in Escherichia coli K-12. HU inhibits ribonucleotide reductase (RNR), an enzyme that catalyzes the formation of deoxyribonucleotides. Unexpectedly, seven of the mutants lacked genes that are required for the incorporation of sulfur into a specific tRNA modification base, 5-methylaminomethyl-2-thiouridine (mnm 5 s 2 U), via persulfide relay. We found that the expression of RNR in the mutants was reduced to about one-third both in the absence and presence of HU, while sufficient deoxynucleoside triphosphate (dNTP) was maintained in the mutants in the absence of HU but a shortage occurred in the presence of HU. Trans -supply of an RNR R2 subunit rescued the HU sensitivity of these mutants. The mutants showed high intracellular ATP/ADP ratios, and overexpression of Hda, which catalyzes the conversion of DnaA-ATP to DnaA-ADP, rescued the HU sensitivity of the mutants, suggesting that DnaA-ATP represses RNR expression. The high intracellular ATP/ADP ratios were due to high respiration activity in the mutants. Our data suggested that intracellular redox was inclined toward the reduced state in these mutants, which may explain a change in RNR activity by reduction of the catalytically formed disulfide bond and high respiration activity by the NADH reducing potential. The relation between persulfide relay and intracellular redox is discussed.
    Type of Medium: Online Resource
    ISSN: 0021-9193 , 1098-5530
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2013
    detail.hit.zdb_id: 1481988-0
    SSG: 12
    Location Call Number Limitation Availability
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  • 2
    Online Resource
    Online Resource
    American Society for Microbiology ; 2013
    In:  Journal of Bacteriology Vol. 195, No. 16 ( 2013-08-15), p. 3796-3796
    In: Journal of Bacteriology, American Society for Microbiology, Vol. 195, No. 16 ( 2013-08-15), p. 3796-3796
    Type of Medium: Online Resource
    ISSN: 0021-9193 , 1098-5530
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 2013
    detail.hit.zdb_id: 1481988-0
    SSG: 12
    Location Call Number Limitation Availability
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  • 3
    Online Resource
    Online Resource
    American Society for Microbiology ; 1998
    In:  Journal of Bacteriology Vol. 180, No. 11 ( 1998-06), p. 2931-2935
    In: Journal of Bacteriology, American Society for Microbiology, Vol. 180, No. 11 ( 1998-06), p. 2931-2935
    Abstract: Escherichia coli has only a single copy of a gene for tRNA 6 Leu (Y. Komine et al., J. Mol. Biol. 212:579–598, 1990). The anticodon of this tRNA is CAA (the wobble position C is modified to O 2 -methylcytidine), and it recognizes the codon UUG. Since UUG is also recognized by tRNA 4 Leu , which has UAA (the wobble position U is modified to 5-carboxymethylaminomethyl- O 2 -methyluridine) as its anticodon, tRNA 6 Leu is not essential for protein synthesis. The BT63 strain has a mutation in the anticodon of tRNA 6 Leu with a change from CAA to CUA, which results in the amber suppressor activity of this strain ( supP , Su + 6). We isolated 18 temperature-sensitive (ts) mutants of the BT63 strain whose temperature sensitivity was complemented by introduction of the wild-type gene for tRNA 6 Leu . These tRNA 6 Leu -requiring mutants were classified into two groups. The 10 group I mutants had a mutation in the miaA gene, whose product is involved in a modification of tRNAs that stabilizes codon-anticodon interactions. Overexpression of the gene for tRNA 4 Leu restored the growth of group I mutants at 42°C. Replacement of the CUG codon with UUG reduced the efficiency of translation in group I mutants. These results suggest that unmodified tRNA 4 Leu poorly recognizes the UUG codon at 42°C and that the wild-type tRNA 6 Leu is required for translation in order to maintain cell viability. The mutations in the six group II mutants were complemented by introduction of the gidA gene, which may be involved in cell division. The reduced efficiency of translation caused by replacement of the CUG codon with UUG was also observed in group II mutants. The mechanism of requirement for tRNA 6 Leu remains to be investigated.
    Type of Medium: Online Resource
    ISSN: 0021-9193 , 1098-5530
    Language: English
    Publisher: American Society for Microbiology
    Publication Date: 1998
    detail.hit.zdb_id: 1481988-0
    SSG: 12
    Location Call Number Limitation Availability
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