Alterations in the solubility and intracellular localization of parkin by several familial Parkinson's disease‐linked point mutations

Mutations in the parkin gene, which encodes a ubiquitin ligase, are currently recognized as the main contributor to familial forms of Parkinson's disease (PD). A simple assumption about the effects of PD‐linked mutations in parkin is that they impair or ablate the enzyme activity. However, a number of recent studies, including ours, have indicated that many disease‐linked point mutants of parkin retain substantial catalytic activity. To understand how the plethora of mutations on parkin contribute to its dysfunction, we have conducted a systematic analysis of a significant number of parkin point mutants (22 in total), which represent the majority of parkin missense/nonsense mutations reported to date. We found that more than half of these mutations, including many located outside of the parkin RING fingers, produce alteration in the solubility of parkin which influences its detergent extraction property. This mutation‐mediated alteration in parkin solubility is also associated with its propensity to form intracellular, aggresome‐like, protein aggregates. However, they do not represent sites where parkin substrates become sequestered. As protein aggregation sequesters the functional forms away from their normal sites of action, our results suggest that alterations in parkin solubility and intracellular localization may underlie the molecular basis of the loss of function caused by several of its mutations.

[1]  S. Shin,et al.  Formation and removal of alpha-synuclein aggregates in cells exposed to mitochondrial inhibitors. , 2002, The Journal of biological chemistry.

[2]  T. Dawson,et al.  Molecular Pathways of Neurodegeneration in Parkinson's Disease , 2003, Science.

[3]  N. Hattori,et al.  An Unfolded Putative Transmembrane Polypeptide, which Can Lead to Endoplasmic Reticulum Stress, Is a Substrate of Parkin , 2001, Cell.

[4]  T. Dawson,et al.  Parkin functions as an E2-dependent ubiquitin- protein ligase and promotes the degradation of the synaptic vesicle-associated protein, CDCrel-1. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[5]  John Q. Trojanowski,et al.  Induction of α-Synuclein Aggregation by Intracellular Nitrative Insult , 2001, The Journal of Neuroscience.

[6]  A. Abeliovich,et al.  Parkin Is a Component of an SCF-like Ubiquitin Ligase Complex and Protects Postmitotic Neurons from Kainate Excitotoxicity , 2003, Neuron.

[7]  R. Kopito,et al.  Aggresomes, inclusion bodies and protein aggregation. , 2000, Trends in cell biology.

[8]  C. Ross,et al.  Parkin ubiquitinates the α-synuclein–interacting protein, synphilin-1: implications for Lewy-body formation in Parkinson disease , 2001, Nature Medicine.

[9]  M. Mouradian,et al.  Parkin Accumulation in Aggresomes Due to Proteasome Impairment* , 2002, The Journal of Biological Chemistry.

[10]  P. Lockhart,et al.  Parkin genetics: one model for Parkinson's disease. , 2004, Human molecular genetics.

[11]  C A Ross,et al.  Synphilin‐1 is present in Lewy bodies in Parkinson's disease , 2000, Annals of neurology.

[12]  K. Lim,et al.  Molecular Mechanisms of Neurodegeneration in Parkinson's Disease: Clues from Mendelian Syndromes , 2003, IUBMB life.

[13]  C. Pickart,et al.  Mechanisms underlying ubiquitination. , 2001, Annual review of biochemistry.

[14]  G. Scott,et al.  Inhibition of proteasomal activity causes inclusion formation in neuronal and non-neuronal cells overexpressing Parkin. , 2003, Molecular biology of the cell.

[15]  L. Seeberger,et al.  Heterozygosity for a mutation in the parkin gene leads to later onset Parkinson disease , 2003, Neurology.

[16]  J. Trojanowski,et al.  Induction of alpha-synuclein aggregation by intracellular nitrative insult. , 2001, The Journal of neuroscience : the official journal of the Society for Neuroscience.

[17]  S. Minoshima,et al.  Mutations in the parkin gene cause autosomal recessive juvenile parkinsonism , 1998, Nature.

[18]  Bonifati,et al.  Association between early-onset Parkinson's disease and mutations in the parkin gene. , 2000, The New England journal of medicine.

[19]  Nobutaka Hattori,et al.  Ubiquitination of a New Form of α-Synuclein by Parkin from Human Brain: Implications for Parkinson's Disease , 2001, Science.

[20]  M. Ruberg,et al.  The C289G and C418R missense mutations cause rapid sequestration of human Parkin into insoluble aggregates , 2003, Neurobiology of Disease.

[21]  P. Bork,et al.  A novel transactivation domain in parkin. , 1999, Trends in biochemical sciences.

[22]  Jian Feng,et al.  Parkin is recruited to the centrosome in response to inhibition of proteasomes , 2003, Journal of Cell Science.

[23]  Y. Imai,et al.  Parkin Suppresses Unfolded Protein Stress-induced Cell Death through Its E3 Ubiquitin-protein Ligase Activity* , 2000, The Journal of Biological Chemistry.

[24]  P. Lockhart,et al.  RING finger 1 mutations in Parkin produce altered localization of the protein. , 2003, Human molecular genetics.

[25]  K. Winklhofer,et al.  Inactivation of Parkin by Oxidative Stress and C-terminal Truncations , 2003, Journal of Biological Chemistry.

[26]  Jean-François Deleuze,et al.  Complex relationship between Parkin mutations and Parkinson disease. , 2002, American journal of medical genetics.

[27]  S. Shin,et al.  Formation and Removal of α-Synuclein Aggregates in Cells Exposed to Mitochondrial Inhibitors* , 2002, The Journal of Biological Chemistry.

[28]  N. Wood,et al.  Parkin is recruited into aggresomes in a stress-specific manner: over-expression of parkin reduces aggresome formation but can be dissociated from parkin's effect on neuronal survival. , 2003, Human molecular genetics.

[29]  Ping Wang,et al.  Structure of a c-Cbl–UbcH7 Complex RING Domain Function in Ubiquitin-Protein Ligases , 2000, Cell.

[30]  L. Pallanck,et al.  Parkin A Multipurpose Neuroprotective Agent? , 2003, Neuron.

[31]  P. Lockhart,et al.  Functional association of the parkin gene promoter with idiopathic Parkinson's disease. , 2002, Human molecular genetics.

[32]  D. Hernandez,et al.  Lewy bodies and parkinsonism in families with parkin mutations , 2001, Annals of neurology.

[33]  Shinsei Minoshima,et al.  Familial Parkinson disease gene product, parkin, is a ubiquitin-protein ligase , 2000, Nature Genetics.