RRC ID |
33573
|
Author |
Hirooka K, Edahiro T, Kimura K, Fujita Y.
|
Title |
Direct and indirect regulation of the ycnKJI operon involved in copper uptake through two transcriptional repressors, YcnK and CsoR, in Bacillus subtilis.
|
Journal |
J Bacteriol
|
Abstract |
Northern blot and primer extension analyses revealed that the ycnKJI operon and the ycnL gene of Bacillus subtilis are transcribed from adjacent promoters that are divergently oriented. The ycnK and ycnJ genes encode a DeoR-type transcriptional regulator and a membrane protein involved in copper uptake, respectively. DNA binding experiments showed that the YcnK protein specifically binds to the ycnK-ycnL intergenic region, including a 16-bp direct repeat that is essential for the high binding affinity of YcnK, and that a copper-specific chelator significantly inhibits YcnK's DNA binding. lacZ reporter analysis showed that the ycnK promoter is induced by copper limitation or ycnK disruption. These results are consistent with YcnK functioning as a copper-responsive repressor that derepresses ycnKJI expression under copper limitation. On the other hand, the ycnL promoter was hardly induced by copper limitation, but ycnK disruption resulted in a slight induction of the ycnL promoter, suggesting that YcnK also represses ycnL weakly. Moreover, while the CsoR protein did not bind to the ycnK-ycnL intergenic region, lacZ reporter analysis demonstrated that csoR disruption induces the ycnK promoter only in the presence of intact ycnK and copZA genes. Since the copZA operon is involved in copper export and repressed by CsoR, it appears that the constitutive copZA expression brought by csoR disruption causes intracellular copper depletion, which releases the repression of the ycnKJI operon by YcnK.
|
Volume |
194(20)
|
Pages |
5675-87
|
Published |
2012-10-1
|
DOI |
10.1128/JB.00919-12
|
PII |
JB.00919-12
|
PMID |
22904286
|
PMC |
PMC3458678
|
MeSH |
Artificial Gene Fusion
Bacillus subtilis / genetics*
Bacillus subtilis / metabolism*
Base Sequence
Blotting, Northern
Copper / metabolism*
DNA, Bacterial / metabolism
Gene Deletion
Gene Expression Profiling
Gene Expression Regulation, Bacterial*
Genes, Reporter
Models, Biological
Molecular Sequence Data
Operon
Promoter Regions, Genetic
Protein Binding
Repressor Proteins / metabolism*
Transcription Factors / metabolism*
Transcription, Genetic*
beta-Galactosidase / analysis
beta-Galactosidase / genetics
|
IF |
3.006
|
Times Cited |
9
|
WOS Category
|
MICROBIOLOGY
|
Resource |
Prokaryotes B. subtilis |
MBS356
MBS357
MBS358
MBS359
MBS360
MBS361
MBS362
MBS363
MBS364
MBS365
MBS366
MBS367
MBS368
MBS369
MBS370 |