TY - JOUR
T1 - Structural basis for tRNA-dependent cysteine biosynthesis
AU - Chen, Meirong
AU - Kato, Koji
AU - Kubo, Yume
AU - Tanaka, Yoshikazu
AU - Liu, Yuchen
AU - Long, Feng
AU - Whitman, William B.
AU - Lill, Pascal
AU - Gatsogiannis, Christos
AU - Raunser, Stefan
AU - Shimizu, Nobutaka
AU - Shinoda, Akira
AU - Nakamura, Akiyoshi
AU - Tanaka, Isao
AU - Yao, Min
N1 - Funding Information:
The synchrotron radiation experiments were performed with the approval of SPring-8 (Proposal No. 2015B1024, 2016A2724) and Photon Factory (Proposal No. 2014G080, 2015R-74, 2016G027, and 2016G141). We thank Ms. Xiaomei Sun for her help in ITC experiment, and Dr. Shinya Saijo for his help in SEC-SAXS experiment and the beamline staff of SPring-8 and Photon Factory for their assistance with data collection. This work was supported by Grant-in-Aid for Scientific Research (B) (15H04334 to I.T., 17H03637 to M.Y.) from the Ministry of Education, Culture, Sports, Science and Technology, Japan, and U.S. National Science Foundation (MCB-1632941 to Y.L. and MCB-1410102 to W.B. W.). S.R. thanks the Max Planck Society and the European Council under the European Union Seventh Framework Programme (FP7/2007–2013) (Grant No. 615984) for supporting this work. M.C. was supported by the International Graduate Program (IGP) ‘Training Program for Global Leaders in Life Science’.
Publisher Copyright:
© 2017 The Author(s).
PY - 2017/12/1
Y1 - 2017/12/1
N2 - Cysteine can be synthesized by tRNA-dependent mechanism using a two-step indirect pathway, where O-phosphoseryl-tRNA synthetase (SepRS) catalyzes the ligation of a mismatching O-phosphoserine (Sep) to tRNACys followed by the conversion of tRNA-bounded Sep into cysteine by Sep-tRNA:Cys-tRNA synthase (SepCysS). In ancestral methanogens, a third protein SepCysE forms a bridge between the two enzymes to create a ternary complex named the transsulfursome. By combination of X-ray crystallography, SAXS and EM, together with biochemical evidences, here we show that the three domains of SepCysE each bind SepRS, SepCysS, and tRNACys, respectively, which mediates the dynamic architecture of the transsulfursome and thus enables a global long-range channeling of tRNACys between SepRS and SepCysS distant active sites. This channeling mechanism could facilitate the consecutive reactions of the two-step indirect pathway of Cys-tRNACys synthesis (tRNA-dependent cysteine biosynthesis) to prevent challenge of translational fidelity, and may reflect the mechanism that cysteine was originally added into genetic code.
AB - Cysteine can be synthesized by tRNA-dependent mechanism using a two-step indirect pathway, where O-phosphoseryl-tRNA synthetase (SepRS) catalyzes the ligation of a mismatching O-phosphoserine (Sep) to tRNACys followed by the conversion of tRNA-bounded Sep into cysteine by Sep-tRNA:Cys-tRNA synthase (SepCysS). In ancestral methanogens, a third protein SepCysE forms a bridge between the two enzymes to create a ternary complex named the transsulfursome. By combination of X-ray crystallography, SAXS and EM, together with biochemical evidences, here we show that the three domains of SepCysE each bind SepRS, SepCysS, and tRNACys, respectively, which mediates the dynamic architecture of the transsulfursome and thus enables a global long-range channeling of tRNACys between SepRS and SepCysS distant active sites. This channeling mechanism could facilitate the consecutive reactions of the two-step indirect pathway of Cys-tRNACys synthesis (tRNA-dependent cysteine biosynthesis) to prevent challenge of translational fidelity, and may reflect the mechanism that cysteine was originally added into genetic code.
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U2 - 10.1038/s41467-017-01543-y
DO - 10.1038/s41467-017-01543-y
M3 - Article
C2 - 29142195
AN - SCOPUS:85034435398
VL - 8
JO - Nature Communications
JF - Nature Communications
SN - 2041-1723
IS - 1
M1 - 1521
ER -