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PMID: 9393713 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Cloning and genetic analysis of the UV resistance determinant (uvr) encoded on the Enterococcus faecalis pheromone-responsive conjugative plasmid pAD1.

Journal of bacteriology ·Vol. 179 ·No. 23 ·1997-12-00 ·Pages 7468-75

Ozawa Y, Tanimoto K, Fujimoto S, Tomita H, Ike Y

Abstract

The conjugative pheromone-responsive plasmid pAD1 (59.6 kb) of Enterococcus faecalis encodes a UV resistance determinant (uvr) in addition to the hemolysin-bacteriocin determinant. pAD1 enhances the UV resistance of wild-type E. faecalis FA2-2 and E. faecalis UV202, which is a UV-sensitive derivative of E. faecalis JH2-2. A 2.972-kb fragment cloned from between 27.7 and 30.6 kb of the pAD1 map conferred UV resistance function on UV202. Sequence analysis showed that the cloned fragment contained three open reading frames designated uvrA, uvrB, and uvrC. The uvrA gene is located on the pAD1 map between 28.1 and 29.4 kb. uvrB is located between 30.1 and 30.3 kb, and uvrC is located between 30.4 and 30.6 kb on the pAD1 map. The uvrA, uvrB, and uvrC genes encode sequences of 442, 60, and 74 amino acids, respectively. The deduced amino acid sequence of the uvrA-encoded protein showed 20% homology of the identical residues with the E. coli UmuC protein. Tn917 insertion mutagenesis and deletion mutant analysis of the cloned fragment showed that uvrA conferred UV resistance. A palindromic sequence, 5'-GAACNGTTC-3', which is identical to the consensus sequence found within the putative promoter region of the Bacillus subtilis DNA damage-inducible genes, was located within the promoter region of uvrA. Two uvrA transcripts of different lengths (i.e., 1.54 and 2.14 kb) which terminate at different points downstream of uvrA were detected in UV202 carrying the deletion mutant containing uvrA. The longer transcript, 2.14 kb, was not detected in UV202 carrying the deletion mutant containing both uvrA and uvrB, which suggests that uvrB encodes a terminator for the uvrA transcript. The uvrA transcript was not detected in any significant quantity in UV202 carrying the cloned fragment containing uvrA, uvrB, and uvrC; on the other hand, the 1.54-kb uvrA transcript was detected in the strain exposed to mitomycin C, which suggests that the UvrC protein functions as a regulator of uvrA.

MeSH Terms
Adenosine Triphosphatases/genetics Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Cloning, Molecular DNA Helicases DNA-Binding Proteins/genetics DNA-Directed DNA Polymerase Dose-Response Relationship, Radiation Endodeoxyribonucleases Enterococcus faecalis/genetics,radiation effects Escherichia coli Proteins Genes, Bacterial Molecular Sequence Data Mutagenesis, Insertional Plasmids/genetics RNA, Messenger/analysis Radiation Tolerance/genetics Sequence Analysis, DNA Sequence Deletion Sequence Homology, Amino Acid Ultraviolet Rays
Chemicals
Bacterial Proteins DNA-Binding Proteins Escherichia coli Proteins RNA, Messenger UvrB protein, E coli UmuC protein, E coli DNA-Directed DNA Polymerase Endodeoxyribonucleases UvrC protein, E coli UvrA protein, E coli Adenosine Triphosphatases DNA Helicases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Ozawa Y
Department of Microbiology, Gunma University School of Medicine, Maebashi, Japan.
Tanimoto K
Fujimoto S
Tomita H
Ike Y
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.179.23.7468-7475.1997
Published
1997-12-00
Pages
7468-75
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC179699
Subset
IM
Databases
GENBANK
AB007844
External Links
PubMed source DOI: 10.1128/jb.179.23.7468-7475.1997 PMC Full Text View journal details
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