文献类型：期刊文献

英文题名：CMT2N-causing aminoacylation domain mutants enable Nrp1 interaction with AlaRS

作者：Sun, Litao[1,2];Wei, Na[1];Kuhle, Bernhard[1];Blocquel, David[1];Novick, Scott[3];Matuszek, Zaneta[4];Zhou, Huihao[1,5];He, Weiwei[1,6];Zhang, Jingjing[2];Weber, Thomas[7];Horvath, Rita[8];Latour, Philippe[9];Pan, Tao[4];Schimmel, Paul[1,3];Griffin, Patrick R.[3];Yang, Xiang-Lei[1]

机构：[1]Scripps Res Inst, Dept Mol Med, La Jolla, CA 92037 USA;[2]Sun Yat Sen Univ, Sch Publ Hlth Shenzhen, Guangzhou 510006, Peoples R China;[3]Scripps Res Inst, Dept Mol Med, Jupiter, FL 33458 USA;[4]Univ Chicago, Dept Biochem & Mol Biol, Chicago, IL 60637 USA;[5]Sun Yat Sen Univ, Sch Pharmaceut Sci, Guangzhou 510006, Peoples R China;[6]East China Univ Sci & Technol, Sch Pharm, Shanghai Key Lab New Drug Design, Shanghai 200237, Peoples R China;[7]Dynam Biosensors GmbH, D-82152 Martinsried, Germany;[8]Univ Cambridge, Dept Clin Neurosci, Cambridge CB2 0PY, England;[9]Hosp Civils, Biol & Pathol Dept, F-68500 Lyon, France

年份：2021

卷号：118

期号：13

外文期刊名：PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000637394200016)】；

基金：We thank all the patients and family members who participated in this study for their contribution. Informed consent was obtained from all individuals included in the study. We also thank the staff at beamline 11-1 of Stanford Synchrotron Radiation Lightsource (SSRL) for assistance with X-ray diffraction data collection and the staff at beamline 4-2 for assistance with SAXS data collection. This work was supported by NIH grants GM088278 and NS113583 (to X.-L.Y.), GM125908 (to P.S.), and GM113194 (to T.P.), and by a fellowship from the National Foundation for Cancer Research. L.S. is supported by the National Natural Science Foundation of China (Grant 31971147) and the Shenzhen Science and Technology Innovation Commission (JCYJ20190807155011406). This work was also supported by the European Research Council (309548 to R.H.), the Wellcome Investigator Award (109915/Z/15/Z to R.H.), the Medical Research Council (UK) (MR/N025431/1 to R.H.), and the Newton Fund (UK/Turkey, MR/N027302/1 to R.H.).

语种：英文

外文关键词：Charcot-Marie-Tooth disease; neuropilin 1; AlaRS

摘要：Through dominant mutations, aminoacyl-tRNA synthetases constitute the largest protein family linked to Charcot-Marie-Tooth disease (CMT). An example is CMT subtype 2N (CMT2N), caused by individual mutations spread out in AlaRS, including three in the aminoacylation domain, thereby suggesting a role for a tRNA-charging defect. However, here we found that two are aminoacylation defective but that the most widely distributed R329H is normal as a purified protein in vitro and in unfractionated patient cell samples. Remarkably, in contrast to wild-type (WT) AlaRS, all three mutant proteins gained the ability to interact with neuropilin 1 (Nrp1), the receptor previously linked to CMT pathogenesis in GlyRS. The aberrant AlaRS-Nrp1 interaction is further confirmed in patient samples carrying the R329H mutation. However, CMT2N mutations outside the aminoacylation domain do not induce the Nrp1 interaction. Detailed biochemical and biophysical investigations, including X-ray crystallography, small-angle X-ray scattering, hydrogen-deuterium exchange (HDX), switchSENSE hydrodynamic diameter determinations, and protease digestions reveal a mutationinduced structural loosening of the aminoacylation domain that correlates with the Nrp1 interaction. The b1b2 domains of Nrp1 are responsible for the interaction with R329H AlaRS. The results suggest Nrp1 is more broadly associated with CMT-associated members of the tRNA synthetase family. Moreover, we revealed a distinct structural loosening effect induced by a mutation in the editing domain and a lack of conformational impact with C-Ala domain mutations, indicating mutations in the same protein may cause neuropathy through different mechanisms. Our results show that, as with other CMT-associated tRNA synthetases, aminoacylation per se is not relevant to the pathology.