TY - JOUR
T1 - Insight into human Miro1/2 domain organization based on the structure of its N-terminal GTPase
AU - Smith, Kyle P.
AU - Focia, Pamela J.
AU - Chakravarthy, Srinivas
AU - Landahl, Eric C.
AU - Klosowiak, Julian L.
AU - Rice, Sarah E.
AU - Freymann, Douglas M.
N1 - Funding Information:
We thank Samuel Light and the Anderson lab for crystallization supplies and thoughtful feedback. We thank Melissa Gonzalez for assistance, and thank Theint Aung, Chi-Hao Luan, and David Haselbach for assisting with data collection and interpretation. We thank Priscilla Yeung and Jodi Nunnari for thoughtful discussions. K.P.S. and J.L.K. were supported by T32GM008382 and the Walter S. and Lucienne Driskill Graduate Training Program in Life Sciences at Northwestern University. S.E.R. was supported by R01GM107209. This work used resources of the Northwestern University Structural Biology Facility and the Keck Biophysics Facility, supported by NCI-CCSG-P30-CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract No. DE-AC02-06CH11357. Use of the Pilatus 3 1M detector was provided by grant 1S10OD018090-01 from NIGMS. Use of the LS-CAT was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (085P1000817). This project was supported by grant 9 P41 GM103622 from the National Institute of General Medical Sciences of the National Institutes of Health. The authors declare no competing financial interests.
Funding Information:
We thank Samuel Light and the Anderson lab for crystallization supplies and thoughtful feedback. We thank Melissa Gonzalez for assistance, and thank Theint Aung, Chi-Hao Luan, and David Haselbach for assisting with data collection and interpretation. We thank Priscilla Yeung and Jodi Nunnari for thoughtful discussions. K.P.S. and J.L.K. were supported by T32GM008382 and the Walter S. and Lucienne Driskill Graduate Training Program in Life Sciences at Northwestern University. S.E.R. was supported by R01GM107209. This work used resources of the Northwestern University Structural Biology Facility and the Keck Biophysics Facility, supported by NCI-CCSG-P30-CA060553 awarded to the Robert H. Lurie Comprehensive Cancer Center. Use of the Advanced Photon Source was supported by the US Department of Energy, Office of Science, Office of Basic Energy Sciences, under Contract No. DE-AC02-06CH11357. Use of the Pilatus 3 1M detector was provided by grant 1S10OD018090-01 from NIGMS. Use of the LS-CAT was supported by the Michigan Economic Development Corporation and the Michigan Technology Tri-Corridor (085P1000817). This project was supported by grant 9 P41 GM103622 from the National Institute of General Medical Sciences of the National Institutes of Health. The authors declare no competing financial interests.
Publisher Copyright:
© 2020 Elsevier Inc.
PY - 2020/12/1
Y1 - 2020/12/1
N2 - Dysfunction in mitochondrial dynamics is believed to contribute to a host of neurological disorders and has recently been implicated in cancer metastasis. The outer mitochondrial membrane adapter protein Miro functions in the regulation of mitochondrial mobility and degradation, however, the structural basis for its roles in mitochondrial regulation remain unknown. Here, we report a 1.7Å crystal structure of N-terminal GTPase domain (nGTPase) of human Miro1 bound unexpectedly to GTP, thereby revealing a non-catalytic configuration of the putative GTPase active site. We identify two conserved surfaces of the nGTPase, the “SELFYY” and “ITIP” motifs, that are potentially positioned to mediate dimerization or interaction with binding partners. Additionally, we report small angle X-ray scattering (SAXS) data obtained from the intact soluble HsMiro1 and its paralog HsMiro2. Taken together, the data allow modeling of a crescent-shaped assembly of the soluble domain of HsMiro1/2. PDB rseference: Crystal structure of the human Miro1 N-terminal GTPase bound to GTP, 6D71.
AB - Dysfunction in mitochondrial dynamics is believed to contribute to a host of neurological disorders and has recently been implicated in cancer metastasis. The outer mitochondrial membrane adapter protein Miro functions in the regulation of mitochondrial mobility and degradation, however, the structural basis for its roles in mitochondrial regulation remain unknown. Here, we report a 1.7Å crystal structure of N-terminal GTPase domain (nGTPase) of human Miro1 bound unexpectedly to GTP, thereby revealing a non-catalytic configuration of the putative GTPase active site. We identify two conserved surfaces of the nGTPase, the “SELFYY” and “ITIP” motifs, that are potentially positioned to mediate dimerization or interaction with binding partners. Additionally, we report small angle X-ray scattering (SAXS) data obtained from the intact soluble HsMiro1 and its paralog HsMiro2. Taken together, the data allow modeling of a crescent-shaped assembly of the soluble domain of HsMiro1/2. PDB rseference: Crystal structure of the human Miro1 N-terminal GTPase bound to GTP, 6D71.
KW - Crystal structure
KW - GTP-binding protein
KW - Gem1p
KW - Miro
KW - Mitochondrial dynamics
KW - RhoT
UR - http://www.scopus.com/inward/record.url?scp=85094600595&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=85094600595&partnerID=8YFLogxK
U2 - 10.1016/j.jsb.2020.107656
DO - 10.1016/j.jsb.2020.107656
M3 - Article
C2 - 33132189
AN - SCOPUS:85094600595
SN - 1047-8477
VL - 212
JO - Journal of Structural Biology
JF - Journal of Structural Biology
IS - 3
M1 - 107656
ER -
