TY - JOUR
T1 - Crystallographic analysis of a subcomplex of the transsulfursome with tRNA for Cys-tRNACys synthesis
AU - Chen, Meirong
AU - Nakazawa, Yuto
AU - Kubo, Yume
AU - Asano, Nozomi
AU - Kato, Koji
AU - Tanaka, Isao
AU - Yao, Min
N1 - Funding Information:
We are grateful to the beamline staff of SPring-8 for their help in data collection. MC is supported by the International Graduate Program (IGP) 'Training Program for Global Leaders in Life Science'. This research was supported by a Grant-in-Aid for Scientific Research (B) (No. 15H04334 to IT) from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
Publisher Copyright:
© 2016 International Union of Crystallography.
PY - 2016
Y1 - 2016
N2 - In most organisms, Cys-tRNACys is directly synthesized by cysteinyl-tRNA synthetase (CysRS). Many methanogenic archaea, however, use a two-step, indirect pathway to synthesize Cys-tRNACys owing to a lack of CysRS and cysteine-biosynthesis systems. This reaction is catalyzed by O-phosphoseryl-tRNA synthetase (SepRS), Sep-tRNA:Cys-tRNA synthase (SepCysS) and SepRS/SepCysS pathway enhancer (SepCysE) as the transsulfursome, in which SepCysE connects both SepRS and SepCysS. On the transsulfursome, SepRS first ligates an O-phosphoserine to tRNACys, and the mischarged intermediate Sep-tRNACys is then transferred to SepCysS, where it is further modified to Cys-tRNACys. In this study, a subcomplex of the transsulfursome with tRNACys (SepCysS-SepCysE-tRNACys), which is involved in the second reaction step of the indirect pathway, was constructed and then crystallized. The crystals diffracted X-rays to a resolution of 2.6 Å and belonged to space group P6522, with unit-cell parameters a = b = 107.2, c = 551.1 Å. The structure determined by molecular replacement showed that the complex consists of a SepCysS dimer, a SepCysE dimer and one tRNACys in the asymmetric unit.
AB - In most organisms, Cys-tRNACys is directly synthesized by cysteinyl-tRNA synthetase (CysRS). Many methanogenic archaea, however, use a two-step, indirect pathway to synthesize Cys-tRNACys owing to a lack of CysRS and cysteine-biosynthesis systems. This reaction is catalyzed by O-phosphoseryl-tRNA synthetase (SepRS), Sep-tRNA:Cys-tRNA synthase (SepCysS) and SepRS/SepCysS pathway enhancer (SepCysE) as the transsulfursome, in which SepCysE connects both SepRS and SepCysS. On the transsulfursome, SepRS first ligates an O-phosphoserine to tRNACys, and the mischarged intermediate Sep-tRNACys is then transferred to SepCysS, where it is further modified to Cys-tRNACys. In this study, a subcomplex of the transsulfursome with tRNACys (SepCysS-SepCysE-tRNACys), which is involved in the second reaction step of the indirect pathway, was constructed and then crystallized. The crystals diffracted X-rays to a resolution of 2.6 Å and belonged to space group P6522, with unit-cell parameters a = b = 107.2, c = 551.1 Å. The structure determined by molecular replacement showed that the complex consists of a SepCysS dimer, a SepCysE dimer and one tRNACys in the asymmetric unit.
KW - Cys-tRNA
KW - SepCysE
KW - SepCysS
KW - SepRS
KW - transsulfursome
UR - http://www.scopus.com/inward/record.url?scp=84978039420&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84978039420&partnerID=8YFLogxK
U2 - 10.1107/S2053230X16009559
DO - 10.1107/S2053230X16009559
M3 - Article
C2 - 27380375
AN - SCOPUS:84978039420
SN - 1744-3091
VL - 72
SP - 569
EP - 572
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
ER -