Organization and transcriptional analysis of a six-gene cluster around the rplK-rplA operon of Corynebacterium glutamicum encoding the ribosomal proteins L11 and L1

A cluster of six genes, tRNA(Trp)-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 Nus...

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Bibliographic Details
Published in:Applied and environmental microbiology 2001-05, Vol.67 (5), p.2183-2190
Main Authors: BARREIRO, Carlos, GONZALEZ-LAVADO, Eva, MARTIN, Juan F
Format: Article
Language:English
Subjects:
Bacteria
Bacteriology
Base Sequence
Biological and medical sciences
Blotting, Northern
Cloning, Molecular
Corynebacterium - genetics
Corynebacterium - growth & development
Corynebacterium - metabolism
Corynebacterium glutamicum
Fundamental and applied biological sciences. Psychology
Gene Expression Regulation, Bacterial
Genes
Genes, Bacterial
Genetics
Genetics and Molecular Biology
Genomics
Microbiology
Molecular Sequence Data
Multigene Family
nusG gene
Operon - genetics
pkwR gene
Plasmids - genetics
Promoter Regions, Genetic - genetics
Proteins
Ribonucleic acid
ribosomal protein L1
ribosomal protein L11
Ribosomal Proteins - genetics
Ribosomal Proteins - metabolism
RNA
rplA gene
rplK gene
secE gene
Sequence Analysis, DNA
Transcription, Genetic
tRNA Trp
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Summary:A cluster of six genes, tRNA(Trp)-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.
ISSN:0099-2240
1098-5336
DOI:10.1128/AEM.67.5.2183-2190.2001