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Applied and Environmental Microbiology, May 2001, p. 2183-2190, Vol. 67, No. 5
Instituto de Biotecnologia (INBIOTEC), Parque
Cientifico de León, 24006 León,1 and
Facultad de Ciencias Biológicas y Ambientales, Area de
Microbiologia, Universidad de León, 24071 León,2 Spain
Received 5 December 2000/Accepted 22 February 2001
A cluster of six genes,
tRNATrp-secE-nusG-rplK-rplA-pkwR, was cloned
and sequenced from a Corynebacterium glutamicum cosmid
library and shown to be contiguous in the C. glutamicum
genome. These genes encode a tryptophanyl tRNA, the protein translocase
component SecE, the antiterminator protein NusG, and the ribosomal
proteins L11 and L1 in addition to PkwR, a putative regulatory protein of the LacI-GalR family. S1 nuclease mapping analysis revealed that
nusG and rplK are expressed as separate
transcriptional units and rplK and rplA are
cotranscribed as a single mRNA. A 19-nucleotide inverted repeat that
appears to correspond to a transcriptional terminator was located in
the 3' region downstream from nusG. Northern analysis with
different probes confirmed the S1 mapping results and showed that the
secE-rplA four-gene region gives rise to four transcripts.
secE was transcribed as a 0.5-kb monocistronic mRNA,
nusG formed two transcripts of 1.4 and 1.0 kb from
different initiation sites, and the two ribosomal protein genes
rplK and rplA were cotranscribed as a single
mRNA of 1.6 kb. A consensus L1 protein binding sequence was identified
in the leader region of the rplK-rplA transcript,
suggesting that expression of the rplK-rplA cluster was
regulated by autogenous regulation exerted by the L1 protein at the
translation level. The promoters of the nusG and
rplK-rplA genes were subcloned in a novel corynebacterial promoter-probe vector and shown to confer strong expression of the
reporter gene.
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.5.2183-2190.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
Organization and Transcriptional Analysis of a
Six-Gene Cluster around the rplK-rplA Operon of
Corynebacterium glutamicum Encoding the Ribosomal Proteins
L11 and L1
*
Corresponding author. Mailing address: Instituto de
Biotecnología (INBIOTEC), Parque Científico de
León, Avda. del Real, no. 1, 24006 León, Spain. Phone: (34 987) 210308. Fax: (34 987) 210388. E-mail:
degjmm{at}unileon.es.
Applied and Environmental Microbiology, May 2001, p. 2183-2190, Vol. 67, No. 5
0099-2240/01/$04.00+0 DOI: 10.1128/AEM.67.5.2183-2190.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.
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