Lens Enhancement of Ubiquitin Conjugation Activity Reduces Intracellular Aggregation of V 76 D Mutant c D-Crystallin

METHODS. Wild type (wt) and V76D mutant cD-crystallin were fused to red fluorescence protein (RFP) and expressed in human lens epithelial cells. The cellular distribution of the expressed proteins was compared by fluorescence microscopy. The solubility of wtand V76D-cD-crystallin was determined by cellular fractionation and Western blotting. Wt-cDRFP and V76D-cD-RFP were also cotransfected along with a ubiquitin ligase (CHIP) or a ubiquitin-conjugating enzyme (Ubc5) into cells. Levels of wtand V76D-cD-crystallin, the percentage of transfected cells with aggregates, and aggregate size were quantified and compared among different groups.

[1]  Nadinath B. Nillegoda,et al.  A Network of Ubiquitin Ligases Is Important for the Dynamics of Misfolded Protein Aggregates in Yeast* , 2012, The Journal of Biological Chemistry.

[2]  L. Ding,et al.  Oligomerization with wt αA- and αB-crystallins reduces proteasome-mediated degradation of C-terminally truncated αA-crystallin. , 2012, Investigative ophthalmology & visual science.

[3]  H. Querfurth,et al.  Cross‐functional E3 ligases Parkin and C‐terminus Hsp70‐interacting protein in neurodegenerative disorders , 2012, Journal of neurochemistry.

[4]  R. Gupta,et al.  The Common Modification in αA-Crystallin in the Lens, N101D, Is Associated with Increased Opacity in a Mouse Model* , 2011, Journal of Biological Chemistry.

[5]  P. Sorensen,et al.  Niclosamide Prevents the Formation of Large Ubiquitin-Containing Aggregates Caused by Proteasome Inhibition , 2010, PloS one.

[6]  A. Pande,et al.  Cataract-associated mutant E107A of human γD-crystallin shows increased attraction to α-crystallin and enhanced light scattering , 2010, Proceedings of the National Academy of Sciences of the United States of America.

[7]  J. King,et al.  Ubiquitin proteasome pathway-mediated degradation of proteins: effects due to site-specific substrate deamidation. , 2010, Investigative ophthalmology & visual science.

[8]  Min Jae Lee,et al.  Enhancement of Proteasome Activity by a Small-Molecule Inhibitor of Usp14 , 2010, Nature.

[9]  U. Andley αA-crystallin R49Cneo mutation influences the architecture of lens fiber cell membranes and causes posterior and nuclear cataracts in mice , 2009, BMC Ophthalmology.

[10]  N. Srinivasan,et al.  Visualization of in situ intracellular aggregation of two cataract-associated human gamma-crystallin mutants: lose a tail, lose transparency. , 2008, Investigative ophthalmology & visual science.

[11]  J. Nix,et al.  Interactions between the quality control ubiquitin ligase CHIP and ubiquitin conjugating enzymes , 2008, BMC Structural Biology.

[12]  N. Sharpless,et al.  CHIP Deficiency Decreases Longevity, with Accelerated Aging Phenotypes Accompanied by Altered Protein Quality Control , 2008, Molecular and Cellular Biology.

[13]  J. Corral,et al.  Inhibition of proteasome by bortezomib causes intracellular aggregation of hepatic serpins and increases the latent circulating form of antithrombin , 2008, Laboratory Investigation.

[14]  Catherine Cheng,et al.  GammaD-crystallin associated protein aggregation and lens fiber cell denucleation. , 2007, Investigative ophthalmology & visual science.

[15]  Yizhi Liu,et al.  Cytoprotective effects of proteasome β5 subunit overexpression in lens epithelial cells , 2007, Molecular vision.

[16]  N. Srinivasan,et al.  Mutation causing self-aggregation in human gammaC-crystallin leading to congenital cataract. , 2006, Investigative ophthalmology & visual science.

[17]  P A Pevzner,et al.  Age-related changes in human crystallins determined from comparative analysis of post-translational modifications in young and aged lens: does deamidation contribute to crystallin insolubility? , 2006, Journal of proteome research.

[18]  Ismael Al-Ramahi,et al.  CHIP Protects from the Neurotoxicity of Expanded and Wild-type Ataxin-1 and Promotes Their Ubiquitination and Degradation* , 2006, Journal of Biological Chemistry.

[19]  F. Shang,et al.  Glutathiolation enhances the degradation of gammaC-crystallin in lens and reticulocyte lysates, partially via the ubiquitin-proteasome pathway. , 2006, Investigative ophthalmology & visual science.

[20]  Wen-Lang Lin,et al.  Deletion of the Ubiquitin Ligase CHIP Leads to the Accumulation, But Not the Aggregation, of Both Endogenous Phospho- and Caspase-3-Cleaved Tau Species , 2006, The Journal of Neuroscience.

[21]  P. Evans,et al.  The triage of damaged proteins: degradation by the ubiquitin‐proteasome pathway or repair by molecular chaperones , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[22]  P. Cohen,et al.  Chaperoned ubiquitylation--crystal structures of the CHIP U box E3 ubiquitin ligase and a CHIP-Ubc13-Uev1a complex. , 2005, Molecular cell.

[23]  F. Shang,et al.  Selectivity of the ubiquitin pathway for oxidatively modified proteins: relevance to protein precipitation diseases , 2005, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[24]  L. Segovia,et al.  γN‐crystallin and the evolution of the βγ‐crystallin superfamily in vertebrates , 2005, The FEBS journal.

[25]  Neer Asherie,et al.  Decrease in protein solubility and cataract formation caused by the Pro23 to Thr mutation in human gamma D-crystallin. , 2005, Biochemistry.

[26]  L. Szweda,et al.  Ubiquitin‐dependent lysosomal degradation of the HNE‐modified proteins in lens epithelial cells , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[27]  G. Scott,et al.  UCH‐L1 aggresome formation in response to proteasome impairment indicates a role in inclusion formation in Parkinson's disease , 2004, Journal of neurochemistry.

[28]  C. Ross,et al.  Protein aggregation and neurodegenerative disease , 2004, Nature Medicine.

[29]  V. Uversky,et al.  Conformational constraints for amyloid fibrillation: the importance of being unfolded. , 2004, Biochimica et biophysica acta.

[30]  Christopher M Dobson,et al.  Principles of protein folding, misfolding and aggregation. , 2004, Seminars in cell & developmental biology.

[31]  V. Godfrey,et al.  CHIP activates HSF1 and confers protection against apoptosis and cellular stress , 2003, The EMBO journal.

[32]  Moonhee Lee,et al.  Proteasomal inhibition causes the formation of protein aggregates containing a wide range of proteins, including nitrated proteins , 2003, Journal of neurochemistry.

[33]  Neer Asherie,et al.  High-resolution X-ray crystal structures of human gammaD crystallin (1.25 A) and the R58H mutant (1.15 A) associated with aculeiform cataract. , 2003, Journal of molecular biology.

[34]  Keiji Tanaka,et al.  CHIP: a quality-control E3 ligase collaborating with molecular chaperones. , 2003, The international journal of biochemistry & cell biology.

[35]  K. Nakayama,et al.  U-box proteins as a new family of ubiquitin ligases. , 2003, Biochemical and biophysical research communications.

[36]  F. Shang,et al.  Lens fibers have a fully functional ubiquitin-proteasome pathway. , 2002, Experimental eye research.

[37]  L. Neckers,et al.  Chaperone-dependent E3 ubiquitin ligase CHIP mediates a degradative pathway for c-ErbB2/Neu , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[38]  Helmut Fuchs,et al.  V76D mutation in a conserved gD-crystallin region leads to dominant cataracts in mice , 2002, Mammalian Genome.

[39]  M. Hayden,et al.  Mutant DNA-binding domain of HSF4 is associated with autosomal dominant lamellar and Marner cataract , 2002, Nature Genetics.

[40]  R. Kopito,et al.  A Principal Role for the Proteasome in Endoplasmic Reticulum-associated Degradation of Misfolded Intracellular Cystic Fibrosis Transmembrane Conductance Regulator* , 2002, The Journal of Biological Chemistry.

[41]  A. Ciechanover,et al.  The ubiquitin-proteasome proteolytic pathway: destruction for the sake of construction. , 2002, Physiological reviews.

[42]  Keiji Tanaka,et al.  CHIP is a chaperone‐dependent E3 ligase that ubiquitylates unfolded protein , 2001, EMBO reports.

[43]  U. Andley,et al.  Ubiquitin-activating enzyme (E1) isoforms in lens epithelial cells: origin of translation, E2 specificity and cellular localization determined with novel site-specific antibodies. , 2001, Experimental eye research.

[44]  D. Cyr,et al.  CHIP Is a U-box-dependent E3 Ubiquitin Ligase , 2001, The Journal of Biological Chemistry.

[45]  F. Shang,et al.  Removal of oxidatively damaged proteins from lens cells by the ubiquitin-proteasome pathway. , 2001, Experimental eye research.

[46]  G. Lukács,et al.  Cooh-Terminal Truncations Promote Proteasome-Dependent Degradation of Mature Cystic Fibrosis Transmembrane Conductance Regulator from Post-Golgi Compartments , 2001, The Journal of cell biology.

[47]  J. King,et al.  Crystal cataracts: Human genetic cataract caused by protein crystallization , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[48]  M. Frydman,et al.  A nonsense mutation (W9X) in CRYAA causes autosomal recessive cataract in an inbred Jewish Persian family. , 2000, Investigative ophthalmology & visual science.

[49]  Alexander Varshavsky,et al.  The ubiquitin system. , 1998, Annual review of biochemistry.

[50]  J. Brynda,et al.  Link between a novel human gammaD-crystallin allele and a unique cataract phenotype explained by protein crystallography. , 2000, Human molecular genetics.

[51]  S. Kaushal,et al.  Missense mutations in MIP underlie autosomal dominant ‘polymorphic’ and lamellar cataracts linked to 12q , 2000, Nature Genetics.

[52]  D. Weeks,et al.  A juvenile-onset, progressive cataract locus on chromosome 3q21-q22 is associated with a missense mutation in the beaded filament structural protein-2. , 2000, American journal of human genetics.

[53]  J. Hess,et al.  Autosomal-dominant congenital cataract associated with a deletion mutation in the human beaded filament protein gene BFSP2. , 2000, American journal of human genetics.

[54]  J. King,et al.  Molecular basis of a progressive juvenile-onset hereditary cataract. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[55]  Martin Rechsteiner,et al.  Recognition of the polyubiquitin proteolytic signal , 2000, The EMBO journal.

[56]  S. Bhattacharya,et al.  Lens biology: development and human cataractogenesis. , 1999, Trends in genetics : TIG.

[57]  F. Shang,et al.  Ubiquitin-dependent pathway is up-regulated in differentiating lens cells. , 1999, Experimental eye research.

[58]  K. Lampi,et al.  Age-related changes in human lens crystallins identified by HPLC and mass spectrometry. , 1998, Experimental eye research.

[59]  T. Shearer,et al.  Age-related changes in human lens crystallins identified by two-dimensional electrophoresis and mass spectrometry. , 1998, Experimental eye research.

[60]  R. Ferrell,et al.  A novel homeobox gene PITX3 is mutated in families with autosomal-dominant cataracts and ASMD , 1998, Nature Genetics.

[61]  F. Shang,et al.  Activity of Ubiquitin-dependent Pathway in Response to Oxidative Stress , 1997, The Journal of Biological Chemistry.

[62]  E. K. Hoffman,et al.  Proteasome inhibition enhances the stability of mouse Cu Zn superoxide dismutase with mutations linked to familial amyotrophic lateral sclerosis , 1996, Journal of the Neurological Sciences.

[63]  Satoshi Omura,et al.  Degradation of CFTR by the ubiquitin-proteasome pathway , 1995, Cell.

[64]  L. Tsui,et al.  Gamma-crystallins of the human eye lens: expression analysis of five members of the gene family , 1987, Molecular and cellular biology.

[65]  A. Hershko,et al.  Ubiquitin-activating enzyme. Mechanism and role in protein-ubiquitin conjugation. , 1982, The Journal of biological chemistry.

[66]  L. Ding,et al.  Degradation of C-terminal truncated alpha A-crystallins by the ubiquitin-proteasome pathway. , 2007, Investigative ophthalmology & visual science.

[67]  J. P. Jensen,et al.  Expression, purification, and properties of the Ubc4/5 family of E2 enzymes. , 2005, Methods in enzymology.

[68]  B. Wallace,et al.  The P23T cataract mutation causes loss of solubility of folded gammaD-crystallin. , 2004, Journal of molecular biology.

[69]  Holly McDonough,et al.  CHIP: a link between the chaperone and proteasome systems , 2003, Cell stress & chaperones.

[70]  C. Dobson,et al.  Altered aggregation properties of mutant gamma-crystallins cause inherited cataract. , 2002, The EMBO journal.

[71]  D. Cyr,et al.  The Hsc70 co-chaperone CHIP targets immature CFTR for proteasomal degradation , 2000, Nature Cell Biology.

[72]  H. Palmer,et al.  Age-related decline in ubiquitin conjugation in response to oxidative stress in the lens. , 1997, Experimental eye research.

[73]  M. Delaye,et al.  Eye lens proteins and transparency: from light transmission theory to solution X-ray structural analysis. , 1988, Annual review of biophysics and biophysical chemistry.