Identification of the substrate binding site in the N-terminal TBP-like domain of RNase H3.

Seiko Miyashita
Takashi Tadokoro
Clement Angkawidjaja
Dong-Ju You
Yuichi Koga
Kazufumi Takano
Shigenori Kanaya

Ribonuclease H3 from Bacillus stearothermophilus (Bst-RNase H3) has the N-terminal TBP-like substrate-binding domain. To identify the substrate binding site in this domain, the mutant proteins of the intact protein and isolated N-domain, in which six of the seventeen residues corresponding to those involved in DNA binding of TBP are individually mutated to Ala, were constructed. All of them exhibited decreased enzymatic activities and/or substrate-binding affinities when compared to those of the parent proteins, suggesting that the N-terminal domain of RNase H3 uses the flat surface of the β-sheet for substrate binding as TBP to bind DNA. This domain may greatly change conformation upon substrate binding.

FEBS letters

585

14

2313

7

10.1016/j.febslet.2011.05.064

https://doi.org/10.1016/j.febslet.2011.05.064
https://www.ncbi.nlm.nih.gov/pubmed/21664908
http://www.scopus.com/inward/record.url?eid=2-s2.0-79960361216&partnerID=MN8TOARS
http://orcid.org/0000-0002-7548-7314