Title
Gene regulation of two ferredoxin:NADP+oxidoreductases by the redox-responsive regulator SurR in Thermococcus kodakarensis
Author
Ryota Hidese, Keita Yamashita, Kohei Kawazuma, Tamotsu Kanai, Haruyuki Atomi, Tadayuki Imanaka, Shinsuke Fujiwara
Year
2017
Journal
Extremophiles
Abstract
The redox-responsive regulator SurR in the hyperthermophilic archaea Pyrococcus furiosusand Thermococcus kodakarensis binds to the SurR-binding consensus sequence (SBS) by responding to the presence of elemental sulfur. Here we constructed a surR gene disruption strain (DTS) in T. kodakarensis, and identified the genes that were under SurR control by comparing the transcriptomes of DTS and parent strains. Among these genes, transcript levels of ferredoxin:NADP+ oxidoreductases 1 and 2 (FNOR1 and FNOR2) genes displayed opposite responses to surR deletion, indicating that SurR repressed FNOR1 transcription while enhancing FNOR2 transcription. Each promoter region contains an SBS upstream (uSBS) and downstream (dSBS) of TATA. In addition to in vitro binding assays, we examined the roles of each SBS in vivo. In FNOR1, mutations in either one of the SBSs resulted in a complete loss of repression, indicating that the presence of both SBSs was essential for repression. In FNOR2, uSBS indeed functioned to enhance gene expression, whereas dSBS functioned in gene repression. SurR bound to uSBS2 of FNOR2 more efficiently than to dSBS2 in vitro, which may explain why SurR overall enhances FNOR2 transcription. Further analyses indicated the importance in the distance between uSBS and TATA for transcriptional activation in FNOR2.
Instrument
J-820
Keywords
Circular dichroism, Secondary structure, Biochemistry