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PMID: 8253666 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Isolation of the gene (miaE) encoding the hydroxylase involved in the synthesis of 2-methylthio-cis-ribozeatin in tRNA of Salmonella typhimurium and characterization of mutants.

Journal of bacteriology ·Vol. 175 ·No. 24 ·1993-12-00 ·Pages 7776-85

Persson BC, Björk GR

Abstract

The modified nucleoside 2-methylthio-N-6-isopentenyl adenosine (ms2i6A) is present at position 37 (3' of the anticodon) of tRNAs that read codons beginning with U except tRNA(I,V Ser) in Escherichia coli. Salmonella typhimurium 2-methylthio-cis-ribozeatin (ms2io6A) is found in tRNA, probably in the corresponding species that have ms2i6A in E. coli. The gene (miaE) for the tRNA(ms2io6A)hydroxylase of S. typhimurium was isolated by complementation in E. coli. The miaE gene was localized close to the argI gene at min 99 of the S. typhimurium chromosomal map. Its DNA sequence and transcription pattern together with complementation studies revealed that the miaE gene is the second gene of a dicistronic operon. Southern blot analysis showed that the miaE gene is absent in E. coli, a finding consistent with the absence of the hydroxylated derivative of ms2i6A in this species. Mutants of S. typhimurium which have MudJ inserted in the miaE gene and which, consequently, are blocked in the ms2i6A hydroxylation reaction were isolated. Unexpectedly, such mutants cannot utilize the citric acid cycle intermediates malate, fumarate, and succinate as carbon sources.

Related Genes
MudJ argI miaA miaB miaC miaE supF30
MeSH Terms
Amino Acid Sequence Bacterial Proteins/biosynthesis,genetics Base Sequence Blotting, Northern Chromatography, High Pressure Liquid Codon DNA Primers Escherichia coli/enzymology,genetics Genes, Bacterial Isopentenyladenosine/analogs & derivatives,biosynthesis Mixed Function Oxygenases/biosynthesis,genetics Molecular Sequence Data Mutagenesis Open Reading Frames Operon Plasmids Protein Biosynthesis RNA, Transfer/biosynthesis Salmonella typhimurium/enzymology,genetics Species Specificity Terminator Regions, Genetic Transcription, Genetic
Chemicals
Bacterial Proteins Codon DNA Primers N(6)-(4-hydroxyisopentenyl)-2-methylthioadenosine Isopentenyladenosine RNA, Transfer 2-methylthio-N6-isopentenyl-adenosine(37)-tRNA monooxygenase, Salmonella typhimurium Mixed Function Oxygenases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Persson B C
Department of Microbiology, Umeå University, Sweden.
Björk G R
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.175.24.7776-7785.1993
Published
1993-12-00
Pages
7776-85
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC206952
Subset
IM
Databases
GENBANK
X73368
External Links
PubMed source DOI: 10.1128/jb.175.24.7776-7785.1993 PMC Full Text View journal details
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