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PMID: 1349602 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Cloning, sequencing, and molecular analysis of the groESL operon of Clostridium acetobutylicum.

Journal of bacteriology ·Vol. 174 ·No. 10 ·1992-05-00 ·Pages 3282-9

Narberhaus F, Bahl H

Abstract

The groESL operon of Clostridium acetobutylicum was cloned in Escherichia coli by using a gene probe of E. coli groESL. Sequencing of a positively reacting 2.2-kbp HindIII fragment contained in the recombinant plasmid pFN1 and a 2.5-kbp XbaI fragment present in pFN4 revealed that both fragments partially overlapped and together spanned 3,493 bp of the clostridial chromosome. Two complete open reading frames (288 and 1632 bp) were found and identified as the groES- and groEL-homologous genes of C. acetobutylicum, respectively. The 3' end of a third gene (orfZ), which was divergently transcribed, showed no significant homology to other sequences available in the EMBL and GenBank data bases. The length of the groESL-specific mRNA (2.2 kb), a transcription terminator downstream of groEL, and a transcription start site upstream of groES, identified by primer extension analysis, indicated that groES and groEL of C. acetobutylicum are organized in a bicistronic operon. From the transcription start site, the promoter structure 5'-TTGCTA (17 bp) TATTAT that shows high homology to the consensus promoter sequence of gram-positive bacteria as well as E. coli was deduced. Transcription of the groESL operon was strongly heat inducible, and maximum levels of mRNA were detected 15 min after heat shock from 30 to 42 degrees C. An 11-bp inverted repeat, located between promoter and translation start sites of groES and partially identical with similar structures in front of several heat shock genes of other bacteria, may play an important role in the regulation of heat shock gene expression in this organism.

MeSH Terms
Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Chaperonin 10 Chaperonin 60 Cloning, Molecular Clostridium/genetics Consensus Sequence Escherichia coli/genetics Gene Expression Regulation, Bacterial Heat-Shock Proteins/genetics Molecular Sequence Data Nucleic Acid Conformation Operon/genetics RNA, Messenger/analysis Regulatory Sequences, Nucleic Acid/genetics Repetitive Sequences, Nucleic Acid/genetics Sequence Homology, Nucleic Acid Transcription, Genetic
Chemicals
Bacterial Proteins Chaperonin 10 Chaperonin 60 Heat-Shock Proteins RNA, Messenger
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Narberhaus F
Institut für Mikrobiologie, Georg-August-Universität Göttingen, Germany.
Bahl H
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.174.10.3282-3289.1992
Published
1992-05-00
Pages
3282-9
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC205997
Subset
IM
Databases
GENBANK
M74572, M79367, M79368, M79369, M79370, M79371, M79372, M87491, M87492, M87836
External Links
PubMed source DOI: 10.1128/jb.174.10.3282-3289.1992 PMC Full Text View journal details
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