TY - JOUR
AU - Halawani, Dalia
AU - Gogonea, Valentin
AU - DiDonato, Joseph A.
AU - Pipich, Vitaliy
AU - Yao, Peng
AU - China, Arnab
AU - Topbas, Celalettin
AU - Vasu, Kommireddy
AU - Arif, Abul
AU - Hazen, Stanley L.
AU - Fox, Paul L.
TI - Structural control of caspase-generated glutamyl-tRNA synthetase by appended noncatalytic WHEP domains
JO - The journal of biological chemistry
VL - 293
IS - 23
SN - 1083-351X
CY - Bethesda, Md.
PB - Soc.72889
M1 - FZJ-2018-06072
SP - 8843 - 8860
PY - 2018
AB - Aminoacyl-tRNA synthetases are ubiquitous, evolutionarilyconserved enzymes catalyzing the conjugation of amino acidsonto cognate tRNAs. During eukaryotic evolution, tRNA syn-thetases have been the targets of persistent structural modifica-tions. These modifications can be additive, as in the evolution-ary acquisition of noncatalytic domains, or subtractive, as in thegeneration of truncated variants through regulated mechanismssuch as proteolytic processing, alternative splicing, or codingregion polyadenylation. A unique variant is the human glu-tamyl-prolyl-tRNA synthetase (EPRS) consisting of two fusedsynthetases joined by a linker containing three copies of theWHEP domain (termed by its presence in tryptophanyl-, histi-dyl-, and glutamyl-prolyl-tRNA synthetases). Here, we identifysite-selective proteolysis as a mechanism that severs the linkagebetween the EPRS synthetases in vitro and in vivo. Caspaseaction targeted Asp-929 in the third WHEP domain, therebyseparating the two synthetases. Using a neoepitope antibodydirected against the newly exposed C terminus, we demonstrateEPRS cleavage at Asp-929 in vitro and in vivo. Biochemical andbiophysical characterizations of the N-terminally generatedEPRS proteoform containing the glutamyl-tRNA synthetaseand most of the linker, including two WHEP domains, com-bined with structural analysis by small-angle neutron scattering,revealed a role for the WHEP domains in modulating conforma-tions of the catalytic core and GSH–S-transferase–C-terminal-like (GST-C) domain. WHEP-driven conformational rearrange-ment altered GST–C domain interactions and conferreddistinct oligomeric states in solution. Collectively, our resultsreveal long-range conformational changes imposed by theWHEP domains and illustrate how noncatalytic domains canmodulate the global structure of tRNA synthetases in complexeukaryotic systems.
LB - PUB:(DE-HGF)16
C6 - pmid:29643180
UR - <Go to ISI:>//WOS:000434941900010
DO - DOI:10.1074/jbc.M117.807503
UR - https://juser.fz-juelich.de/record/856721
ER -