TY - JOUR
T1 - Human RIPK3 maintains MLKL in an inactive conformation prior to cell death by necroptosis
AU - Meng, Yanxiang
AU - Davies, Katherine A.
AU - Fitzgibbon, Cheree
AU - Young, Samuel N.
AU - Garnish, Sarah E.
AU - Horne, Christopher R.
AU - Luo, Cindy
AU - Garnier, Jean Marc
AU - Liang, Lung Yu
AU - Cowan, Angus D.
AU - Samson, Andre L.
AU - Lessene, Guillaume
AU - Sandow, Jarrod J.
AU - Czabotar, Peter E.
AU - Murphy, James M.
N1 - Funding Information:
We thank Dr. Janet Newman and the C3 Facility for assistance with screening for protein crystallization conditions, and the Australian Synchrotron MX and SAXS/WAXS beamline staff for assistance with data collection. This research was undertaken using the MX2 crystallography beamline at the Australian Synchrotron, Victoria, Australia, and made use of the ACRF Detector. We acknowledge scholarship support for Y.M. (Melbourne Research Scholarship; AINSE PGRA scholarship) and S.E.G. (Australian Government Research Training Program Stipend Scholarship; Wendy Dowsett Scholarship). We are grateful to the National Health and Medical Research Council for fellowship (G.L., 1117089; P.E.C., 1079700; J.M.M., 1172929), grant (A.L.S. and J.J.S., 2002965) and infrastructure (IRIISS 9000653) support; Anaxis Pharma Pty Ltd for funding support; the Australian Cancer Research Foundation; and the Victorian Government Operational Infrastructure Support scheme.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - The ancestral origins of the lytic cell death mode, necroptosis, lie in host defense. However, the dysregulation of necroptosis in inflammatory diseases has led to widespread interest in targeting the pathway therapeutically. This mode of cell death is executed by the terminal effector, the MLKL pseudokinase, which is licensed to kill following phosphorylation by its upstream regulator, RIPK3 kinase. The precise molecular details underlying MLKL activation are still emerging and, intriguingly, appear to mechanistically-diverge between species. Here, we report the structure of the human RIPK3 kinase domain alone and in complex with the MLKL pseudokinase. These structures reveal how human RIPK3 structurally differs from its mouse counterpart, and how human RIPK3 maintains MLKL in an inactive conformation prior to induction of necroptosis. Residues within the RIPK3:MLKL C-lobe interface are crucial to complex assembly and necroptotic signaling in human cells, thereby rationalizing the strict species specificity governing RIPK3 activation of MLKL.
AB - The ancestral origins of the lytic cell death mode, necroptosis, lie in host defense. However, the dysregulation of necroptosis in inflammatory diseases has led to widespread interest in targeting the pathway therapeutically. This mode of cell death is executed by the terminal effector, the MLKL pseudokinase, which is licensed to kill following phosphorylation by its upstream regulator, RIPK3 kinase. The precise molecular details underlying MLKL activation are still emerging and, intriguingly, appear to mechanistically-diverge between species. Here, we report the structure of the human RIPK3 kinase domain alone and in complex with the MLKL pseudokinase. These structures reveal how human RIPK3 structurally differs from its mouse counterpart, and how human RIPK3 maintains MLKL in an inactive conformation prior to induction of necroptosis. Residues within the RIPK3:MLKL C-lobe interface are crucial to complex assembly and necroptotic signaling in human cells, thereby rationalizing the strict species specificity governing RIPK3 activation of MLKL.
UR - https://www.scopus.com/pages/publications/85119894398
U2 - 10.1038/s41467-021-27032-x
DO - 10.1038/s41467-021-27032-x
M3 - Article
C2 - 34811356
AN - SCOPUS:85119894398
SN - 2041-1723
VL - 12
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 6783
ER -
SDG 3 Good Health and Well-being