TY - JOUR
T1 - ATP synthase hexamer assemblies shape cristae of Toxoplasma mitochondria
AU - Mühleip, Alexander
AU - Kock Flygaard, Rasmus
AU - Ovciarikova, Jana
AU - Lacombe, Alice
AU - Fernandes, Paula
AU - Sheiner, Lilach
AU - Amunts, Alexey
N1 - Funding Information:
We acknowledge the ESRF beamline CM01 for provision of beam time for experiment MX-2107, and would like to thank especially Eaazhisai Kandiah for the excellent support. We thank the Astbury Biostructure Laboratory at the University of Leeds for tomography data collection, and especially Rebecca Thompson for her dedicated help. We also thank Michael Hall for data collection at the SciLifeLab National cryo-EM facility. We thank Leandro Lemgruber of Glasgow Imaging Facility for his support and assistance in this work, members of Sheiner lab for help with parasite growth and harvesting, members of the Kuvin Center for the Study of Infectious and Tropical Diseases, Hebrew University of Jerusalem, for providing work space and support with flow cytometry work. This work was funded by the Swedish Foundation for Strategic Research (FFL15:0325), Ragnar Söderberg Foundation (M44/16), European Research Council (ERC-2018-StG-805230), Knut and Alice Wallenberg Foundation (2018.0080), BBSRC (BB/N003675/1), Wellcome Investigator award (217173/Z/19/Z). A.M. is supported by an EMBO Long-Term Fellowship (ALTF 260-2017). A.A. is supported by the EMBO Young Investigator Program. L.S. is a Royal Society of Edinburgh Personal Research Fellow. The SciLifeLab cryo-EM facility is funded by the Knut and Alice Wallenberg, Family Erling Persson, and Kempe foundations.
Publisher Copyright:
© 2021, The Author(s).
Copyright:
Copyright 2020 Elsevier B.V., All rights reserved.
PY - 2021/12/1
Y1 - 2021/12/1
N2 - Mitochondrial ATP synthase plays a key role in inducing membrane curvature to establish cristae. In Apicomplexa causing diseases such as malaria and toxoplasmosis, an unusual cristae morphology has been observed, but its structural basis is unknown. Here, we report that the apicomplexan ATP synthase assembles into cyclic hexamers, essential to shape their distinct cristae. Cryo-EM was used to determine the structure of the hexamer, which is held together by interactions between parasite-specific subunits in the lumenal region. Overall, we identified 17 apicomplexan-specific subunits, and a minimal and nuclear-encoded subunit-a. The hexamer consists of three dimers with an extensive dimer interface that includes bound cardiolipins and the inhibitor IF1. Cryo-ET and subtomogram averaging revealed that hexamers arrange into ~20-megadalton pentagonal pyramids in the curved apical membrane regions. Knockout of the linker protein ATPTG11 resulted in the loss of pentagonal pyramids with concomitant aberrantly shaped cristae. Together, this demonstrates that the unique macromolecular arrangement is critical for the maintenance of cristae morphology in Apicomplexa.
AB - Mitochondrial ATP synthase plays a key role in inducing membrane curvature to establish cristae. In Apicomplexa causing diseases such as malaria and toxoplasmosis, an unusual cristae morphology has been observed, but its structural basis is unknown. Here, we report that the apicomplexan ATP synthase assembles into cyclic hexamers, essential to shape their distinct cristae. Cryo-EM was used to determine the structure of the hexamer, which is held together by interactions between parasite-specific subunits in the lumenal region. Overall, we identified 17 apicomplexan-specific subunits, and a minimal and nuclear-encoded subunit-a. The hexamer consists of three dimers with an extensive dimer interface that includes bound cardiolipins and the inhibitor IF1. Cryo-ET and subtomogram averaging revealed that hexamers arrange into ~20-megadalton pentagonal pyramids in the curved apical membrane regions. Knockout of the linker protein ATPTG11 resulted in the loss of pentagonal pyramids with concomitant aberrantly shaped cristae. Together, this demonstrates that the unique macromolecular arrangement is critical for the maintenance of cristae morphology in Apicomplexa.
KW - Binding Sites
KW - Cardiolipins/chemistry
KW - Cryoelectron Microscopy
KW - Gene Expression
KW - Mitochondria/genetics
KW - Mitochondrial Membranes/metabolism
KW - Mitochondrial Proton-Translocating ATPases/chemistry
KW - Models, Molecular
KW - Protein Binding
KW - Protein Conformation, alpha-Helical
KW - Protein Conformation, beta-Strand
KW - Protein Interaction Domains and Motifs
KW - Protein Multimerization
KW - Protein Subunits/chemistry
KW - Proteins/chemistry
KW - Protozoan Proteins/chemistry
KW - Substrate Specificity
KW - Thermodynamics
KW - Toxoplasma/genetics
UR - http://www.scopus.com/inward/record.url?scp=85098749205&partnerID=8YFLogxK
U2 - 10.1038/s41467-020-20381-z
DO - 10.1038/s41467-020-20381-z
M3 - Journal article
C2 - 33402698
AN - SCOPUS:85098749205
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 120
ER -