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Journal of Bacteriology, December 1999, p. 7256-7265, Vol. 181, No. 23
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Identification of the miaB Gene,
Involved in Methylthiolation of Isopentenylated A37 Derivatives in
the tRNA of Salmonella typhimurium and
Escherichia coli
Birgitta
Esberg,1
Hon-Chiu Eastwood
Leung,2
Ho-Ching
Tiffany
Tsui,2,
Glenn R.
Björk,1,* and
Malcolm E.
Winkler2,
Department of Microbiology, Umeå University,
S-901 87 Umeå, Sweden,1 and Department
of Microbiology and Molecular Genetics, University of Texas, Houston,
Medical School, Houston, Texas 77030-15012
Received 2 June 1999/Accepted 24 September 1999
The tRNA of the miaB2508::Tn10dCm
mutant of Salmonella typhimurium is deficient in the
methylthio group of the modified nucleoside N6-(4-hydroxyisopentenyl)-2-methylthioadenosine
(ms2io6A37). By sequencing, we found that the
Tn10dCm of this strain had been inserted into the
f474 (yleA) open reading frame, which is
located close to the nag locus in both S. typhimurium and Escherichia coli. By complementation
of the miaB2508::Tn10dCm mutation with a
minimal subcloned f474 fragment, we showed that
f474 could be identified as the miaB gene,
which is transcribed in the counterclockwise direction on the bacterial
chromosome. Transcriptional studies revealed two promoters upstream of
miaB in E. coli and S. typhimurium. A Rho-independent terminator was identified downstream of the miaB gene, at which the majority (96%) of the
miaB transcripts terminate in E. coli, showing
that the miaB gene is part of a monocistronic operon. A
highly conserved motif with three cysteine residues was present in
MiaB. This motif resembles iron-binding sites in other proteins. Only a
weak similarity to an AdoMet-binding site was found, favoring the idea
that the MiaB protein is involved in the thiolation step and not in the
methylating reaction of ms2i(o)6A37 formation.
*
Corresponding author. Mailing address: Department of
Microbiology, Umeå University, S-901 87 Umeå, Sweden.
Phone: 46-90-7856756. Fax: 46-90-772630. E-mail:
glenn.bjork{at}micro.umu.se.

Present address: Lilly Research Laboratories, Indianapolis, IN
46285.
Journal of Bacteriology, December 1999, p. 7256-7265, Vol. 181, No. 23
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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