Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity

Capria Rinaldi; Christopher S. Waters; Zizheng Li; Karl Kumbier; Lee Rao; R. Jeremy Nichols; Matthew P. Jacobson; Lani F. Wu; Steven J. Altschuler

Cell Reports (May 2023)

Dissecting the effects of GTPase and kinase domain mutations on LRRK2 endosomal localization and activity

Capria Rinaldi,
Christopher S. Waters,
Zizheng Li,
Karl Kumbier,
Lee Rao,
R. Jeremy Nichols,
Matthew P. Jacobson,
Lani F. Wu,
Steven J. Altschuler

Affiliations

Capria Rinaldi: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
Christopher S. Waters: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
Zizheng Li: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
Karl Kumbier: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
Lee Rao: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
R. Jeremy Nichols: Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA 94305, USA
Matthew P. Jacobson: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA
Lani F. Wu: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA; Corresponding author
Steven J. Altschuler: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA; Corresponding author

Journal volume & issue: Vol. 42, no. 5
p. 112447

Abstract

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Summary: Parkinson’s disease-causing leucine-rich repeat kinase 2 (LRRK2) mutations lead to varying degrees of Rab GTPase hyperphosphorylation. Puzzlingly, LRRK2 GTPase-inactivating mutations—which do not affect intrinsic kinase activity—lead to higher levels of cellular Rab phosphorylation than kinase-activating mutations. Here, we investigate whether mutation-dependent differences in LRRK2 cellular localization could explain this discrepancy. We discover that blocking endosomal maturation leads to the rapid formation of mutant LRRK2+ endosomes on which LRRK2 phosphorylates substrate Rabs. LRRK2+ endosomes are maintained through positive feedback, which mutually reinforces membrane localization of LRRK2 and phosphorylated Rab substrates. Furthermore, across a panel of mutants, cells expressing GTPase-inactivating mutants form strikingly more LRRK2+ endosomes than cells expressing kinase-activating mutants, resulting in higher total cellular levels of phosphorylated Rabs. Our study suggests that the increased probability that LRRK2 GTPase-inactivating mutants are retained on intracellular membranes compared to kinase-activating mutants leads to higher substrate phosphorylation.

CP: Cell biology

Published in Cell Reports

ISSN: 2211-1247 (Online)
Publisher: Elsevier
Country of publisher: Netherlands
LCC subjects: Science: Biology (General)
Website: http://www.cell.com/cell-reports/home

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