期刊
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
卷 389, 期 4, 页码 622-627出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.bbrc.2009.09.038
关键词
Archaea; Hyperthermophile; DEAD-box protein; Cold adaptation; Thermococcus kodakaraensis
资金
- Japan Society for the Promotion of Science (JSPS) [20580090]
- Creative Scientific Research [18GS0421]
- Grants-in-Aid for Scientific Research [20580090] Funding Source: KAKEN
TK0306 (Tk-DeaD) of hyperthermophilic archaeon Thermococcus kodakaraensis is annotated as the DEAD-box helicase gene: nevertheless, its ortholog has not been identified in closely related genera, Pyrococcus spp., which generally grow at higher temperature than T. kodakaraensis, suggesting that the cold-inducible RNA helicase of Tk-DeaD functions under cold stress conditions. Quantitative RT-PCR revealed that Tk-deaD was more dominantly transcribed at 60 degrees C than at 85 degrees C and 93 degrees C in both logarithmic and stationary phases. Immunoblot analyses revealed that Tk-DeaD was detected only in logarithmic-phase cells cultivated at 60 degrees C but hardly detected at 85 degrees C and 93 degrees C in both phases. Tk-DeaD expression is, hence, post-transcriptionally regulated and appears under vigorous growth conditions at 60 degrees C. Recombinant Tk-DeaD purified to homogeneity started to unfold at 20 degrees C, fully unfolded at 70 degrees C, and exhibited maximal ATPase activity and unwinding activity specific for single-strand paired RNA at 50 degrees C, which is lower than the growth limit of T. kodakaraensis. (C) 2009 Elsevier Inc. All rights reserved.
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