Regulation of galactose metabolism through the HisK:GalR two-component system in Thermoanaerobacter tengcongensis.
Thermoanaerobacter tengcongensis could utilize galactose as a carbon source via the enzymes encoded by a novel gal operon, whose regulation mechanism has yet to be elucidated. We propose here that the gal operon in T. tengcongensis is regulated through a HisK:GalR two-component system. By using radioactive isotope assay and genetic analysis, we found that the kinase of this system, HisK, is phosphorylated by ATP, and the regulator, GalR, accepts a phosphoryl group during phosphorelay, in which the phosphoryl group at HisK-His-259 is transferred to GalR-Asp-56. Two-dimensional electrophoresis, followed by Western blotting, revealed that phosphorylation status of GalR is uniquely dependent on the galactose stimulus in vivo. Furthermore, DNA pulldown assays demonstrated that the phosphorylated GalR prefers binding to the operator DNA O(2), whereas the unphosphorylated GalR to O(1). A model of HisK:GalR is proposed to explain how galactose mediates the expression of the gal operon in T. tengcongensis.
Qian
Zhong
Z
Beijing Institute of Genomics, Chinese Academy of Sciences, Beijing Airport Industrial Zone B-6, Shunyi, Beijing, China.
Wang
Quanhui
Q
Tong
Wei
W
Zhou
Chuanqi
C
Wang
Qian
Q
Liu
Siqi
S
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000235:genetics / Q000378:metabolism*; D005690:Galactose / Q000235:genetics / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D000071677:Histidine Kinase; D009876:Operon; D010766:Phosphorylation; D011494:Protein Kinases / Q000235:genetics / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D046968:Thermoanaerobacter / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C050993:Galactose repressor proteins; D012097:Repressor Proteins; D011494:Protein Kinases; D000071677:Histidine Kinase; D005690:Galactose