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

doi:10.1016/j.celrep.2023.112447

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

Cell Rep. 2023 May 30;42(5):112447. doi: 10.1016/j.celrep.2023.112447. Epub 2023 May 3.

Authors

Capria Rinaldi¹, Christopher S Waters¹, Zizheng Li¹, Karl Kumbier¹, Lee Rao¹, R Jeremy Nichols², Matthew P Jacobson¹, Lani F Wu³, Steven J Altschuler⁴

Affiliations

¹ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA.
² Department of Pathology, Stanford University, 300 Pasteur Drive, Stanford, CA 94305, USA.
³ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: lani.wu@ucsf.edu.
⁴ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA 94158, USA. Electronic address: steven.altschuler@ucsf.edu.

PMID: 37141099
DOI: 10.1016/j.celrep.2023.112447

Abstract

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.

Keywords: CP: Cell biology; LRRK2; Parkinson's disease; Rab phosphorylation; VPS34 inhibition; endosomal localization; positive feedback.

Publication types

Research Support, U.S. Gov't, Non-P.H.S.
Research Support, Non-U.S. Gov't

MeSH terms

Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / genetics
Leucine-Rich Repeat Serine-Threonine Protein Kinase-2 / metabolism
Mutation / genetics
Phosphorylation
Protein Serine-Threonine Kinases* / genetics
Protein Serine-Threonine Kinases* / metabolism
rab GTP-Binding Proteins* / genetics
rab GTP-Binding Proteins* / metabolism

Substances

Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
Protein Serine-Threonine Kinases
rab GTP-Binding Proteins