Identification of promoter in the 5'-flanking region of the E. coli thioredoxin-linked thiol peroxidase gene: evidence for the existence of oxygen-related transcriptional regulatory protein.
E. coli thiol peroxidase (Tpx) linked to the thioredoxin as an in vivo thiol regenerating system acts as an antioxidant enzyme removing peroxides and H2O2. In order to elucidate the mechanism regulating tpx gene expression in E. coli in response to oxygen stress, we made 5' progressive deletions of upstream region from tpx gene, and fused to lacZ gene. LacZ activity was increased 6-fold by oxygen stress and inverted repeat sequence located between -47 and -33 nt was proven to be essential for the oxygen response of tpx promoter. Primer extension experiment and analysis of upstream sequence revealed transcription start point, -10, and -35 regions, which are in good agreements with the consensus sequences recognized by E sigma 70. Northern hybridization showed that expression of tpx gene is regulated at the transcriptional level. DNA binding assays using inverted repeat sequence including -35 region provides preliminary evidence that expression of tpx requires additional transcriptional factor in response to oxygen stress.
Kim
H K
HK
Department of Genetic Engineering, Pai Chai University, Seo Gu, Taejon, Republic of Korea.
Kim
S J
SJ
Lee
J W
JW
Lee
J W
JW
Cha
M K
MK
Kim
I H
IH
D000595:Amino Acid Sequence; D001483:Base Sequence; D004247:DNA; D004268:DNA-Binding Proteins / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D029968:Escherichia coli Proteins; D008969:Molecular Sequence Data; D033903:Periplasmic Proteins; D010544:Peroxidases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D017384:Sequence Deletion; D013879:Thioredoxins / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics*
D004268:DNA-Binding Proteins; D029968:Escherichia coli Proteins; D033903:Periplasmic Proteins; D014157:Transcription Factors; D013879:Thioredoxins; D004247:DNA; D010544:Peroxidases; C097049:Tpx protein, E coli