title: The G2385R variant of leucine-rich repeat kinase 2 associated with Parkinson's disease is a partial loss-of-function mutation.
abstract: Autosomal-dominant missense mutations in LRRK2 (leucine-rich repeat kinase 2) are a common genetic cause of PD (Parkinson's disease). LRRK2 is a multidomain protein with kinase and GTPase activities. Dominant mutations are found in the domains that have these two enzyme activities, including the common G2019S mutation that increases kinase activity 2-3-fold. However, there is also a genetic variant in some populations, G2385R, that lies in a C-terminal WD40 domain of LRRK2 and acts as a risk factor for PD. In the present study we show that the G2385R mutation causes a partial loss of the kinase function of LRRK2 and deletion of the C-terminus completely abolishes kinase activity. This effect is strong enough to overcome the kinase-activating effects of the G2019S mutation in the kinase domain. Hsp90 (heat-shock protein of 90 kDa) has an increased affinity for the G2385R variant compared with WT (wild-type) LRRK2, and inhibition of the chaperone binding combined with proteasome inhibition leads to association of mutant LRRK2 with high molecular mass native fractions that probably represent proteasome degradation pathways. The loss-of-function of G2385R correlates with several cellular phenotypes that have been proposed to be kinase-dependent. These results suggest that the C-terminus of LRRK2 plays an important role in maintaining enzymatic function of the protein and that G2385R may be associated with PD in a way that is different from kinase-activating mutations. These results may be important in understanding the differing mechanism(s) by which mutations in LRRK2 act and may also have implications for therapeutic strategies for PD.
authors.affiliation: Cell Biology and Gene Expression Section, National Institute on Aging, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Iakov N
authors.initials: IN
authors.lastname: Rudenko
authors.forename: Alice
authors.initials: A
authors.lastname: Kaganovich
authors.forename: David N
authors.initials: DN
authors.lastname: Hauser
authors.forename: Aleksandra
authors.initials: A
authors.lastname: Beylina
authors.forename: Ruth
authors.initials: R
authors.lastname: Chia
authors.forename: Jinhui
authors.initials: J
authors.lastname: Ding
authors.forename: Dragan
authors.initials: D
authors.lastname: Maric
authors.forename: Howard
authors.initials: H
authors.lastname: Jaffe
authors.forename: Mark R
authors.initials: MR
authors.lastname: Cookson
mesh_terms: D019943:Amino Acid Substitution; D002460:Cell Line / Q000201:enzymology / Q000648:ultrastructure; D002850:Chromatography, Gel; D018841:HSP90 Heat-Shock Proteins / Q000378:metabolism; D006801:Humans; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D008861:Microsomes / Q000378:metabolism; D009154:Mutation; D020125:Mutation, Missense; D016501:Neurites / Q000378:metabolism; D010300:Parkinson Disease / Q000201:enzymology* / Q000235:genetics / Q000503:physiopathology; D010766:Phosphorylation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*
chemical_list: D018841:HSP90 Heat-Shock Proteins; C495280:LRRK2 protein, human; D000071158:Leucine-Rich Repeat Serine-Threonine Protein Kinase-2; D017346:Protein Serine-Threonine Kinases