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Regulatory Characteristics of Bacillus pumilus Protease Promoters
Expression of extracellular protease genes of Bacilli is subject to regulation by many positive and negative regulators. Here we analyzed 5′ regulatory regions of genes encoding proteolytic proteases AprBp, GseBp, and MprBp from Bacillus pumilus strain 3–19. Gfp fusion constructs with upstream genom...
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Published in: | Current microbiology 2017-05, Vol.74 (5), p.550-559 |
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Main Authors: | , , |
Format: | Article |
Language: | English |
Subjects: | |
Citations: | Items that this one cites Items that cite this one |
Online Access: | Get full text |
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Summary: | Expression of extracellular protease genes of
Bacilli
is subject to regulation by many positive and negative regulators. Here we analyzed 5′ regulatory regions of genes encoding proteolytic proteases AprBp, GseBp, and MprBp from
Bacillus pumilus
strain 3–19. Gfp fusion constructs with upstream genomic regions of different lengths were created for all three genes to identify their natural promoters (regulatory regions). Our results suggest that the
aprBp
gene, encoding the major subtilisin-like protease, has the most extensive promoter region of approximately 445 bp, while the minor protease genes encoding glutamyl endopeptidase (
gseBp
) and metalloproteinase (
mprBp
) are preceded by promoters of 150 and 250 bp in length, respectively. Promoter analysis of P
aprBp
-gfpmu3
and P
gseBp
-
gfpmu3
reporter fusion constructs in
degU
and
spo0A
mutants indicates a positive regulatory effect of DegU and Spo0A on protease expression, while the disruption of
abrB, sinR
, and
scoC
repressor genes did not significantly affect promoter activities of all protease genes. On the other hand, the expression of P
aprBp
-gfpmu3
and P
gseBp
-
gfpmu3
reporters increased 1.6- and 3.0-fold, respectively, in
sigD
-deficient cells, indicating that the prevention of motility gene expression promotes protease expression. Our results indicate that all examined regulators regulated serine proteases production in
B. subtilis
. |
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ISSN: | 0343-8651 1432-0991 |
DOI: | 10.1007/s00284-017-1212-3 |