Ubiquitin editing enzyme UCH L1 and microtubule dynamics: Implication in mitosis

Bheda, Anjali; Gullapalli, Anuradha; Caplow, Michael; Pagano, Joseph S.; Shackelford, Julia

doi:10.4161/cc.9.5.10934

Cited by 60 publications

(58 citation statements)

References 46 publications

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“…More recently, a report suggests that UCHL1 can inhibit microtubule formation in a ubiquitinationdependent manner. The authors of this report suggest that UCHL1 may increase ubiquitination (and hence behave as a ligase) of microtubule components (28). In light of these studies, and others that indicate that UCHL1 may have functions independent of the ubiquitin-proteasome system (29,30) and the misaligned active site observed in the structure of the apo form of the protein, the demonstration that UCHL1 can adopt a productive conformation as a hydrolase when bound to ubiquitin assumes particular significance.…”

Section: Al Demonstrating That Leu8 Of Ubiquitin Is Critically Requimentioning

confidence: 94%

Ubiquitin vinyl methyl ester binding orients the misaligned active site of the ubiquitin hydrolase UCHL1 into productive conformation

Boudreaux

Maiti

Davies

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

103

120

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Ubiquitin carboxy-terminal hydrolase L1 (UCHL1) is a Parkinson disease-associated, putative cysteine protease found abundantly and selectively expressed in neurons. The crystal structure of apo UCHL1 showed that the active-site residues are not aligned in a canonical form, with the nucleophilic cysteine being 7.7 Å from the general base histidine, an arrangement consistent with an inactive form of the enzyme. Here we report the crystal structures of the wild type and two Parkinson disease-associated variants of the enzyme, S18Y and I93M, bound to a ubiquitin-based suicide substrate, ubiquitin vinyl methyl ester. These structures reveal that ubiquitin vinyl methyl ester binds primarily at two sites on the enzyme, with its carboxy terminus at the active site and with its amino-terminal β-hairpin at the distal site-a surface-exposed hydrophobic crevice 17 Å away from the active site. Binding at the distal site initiates a cascade of side-chain movements in the enzyme that starts at a highly conserved, surface-exposed phenylalanine and is relayed to the active site resulting in the reorientation and proximal placement of the general base within 4 Å of the catalytic cysteine, an arrangement found in productive cysteine proteases. Mutation of the distal-site, surface-exposed phenylalanine to alanine reduces ubiquitin binding and severely impairs the catalytic activity of the enzyme. These results suggest that the activity of UCHL1 may be regulated by its own substrate.deubiquitinating enzyme | enzyme suicide substrate complex | neurodegeneration | ubiquitination U CHL1, a member of the UCH (ubiquitin C-terminal hydrolase) family of deubiquitinating enzymes (DUBs), is a 223-amino acid protein found abundantly and selectively expressed in brain, constituting up to 1-2% of total brain protein (1, 2). In vivo studies suggest that UCHL1 is involved in regulation of ubiquitin pool, apoptosis, and learning and memory, and its absence in mice because of spontaneous intragenic deletions yields phenotypes with neurological defects (3). Mutations in UCHL1 have been implicated in Parkinson disease (PD). A point mutation near the active site that changes Ile93 to Met (I93M) has been linked to an increased risk of developing an autosomaldominant form of PD (4). Conversely, a common S18Y polymorphism reduces susceptibility to PD (5, 6) and Alzheimer's disease (7). In addition to its association with neurodegenerative diseases, abnormal expression of UCHL1 is found in many forms of cancer, including lung, colorectal, and pancreatic cancers, and may be related to tumor progression (8, 9). The normal function of UCHL1, however, is not known. Also unknown are how the activity of this abundant neuronal enzyme is regulated and what its true physiological substrates are, although biochemical studies have indicated that UCHL1 can accept short-peptide (α or ϵ-amino-linked) or small-molecule C-terminal conjugates of ubiquitin as substrates, cleaving, as its name suggests, the amide bond following immediately after the C-terminal glycine (Gly7...

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Section: Al Demonstrating That Leu8 Of Ubiquitin Is Critically Requimentioning

confidence: 94%

Ubiquitin vinyl methyl ester binding orients the misaligned active site of the ubiquitin hydrolase UCHL1 into productive conformation

Boudreaux

Maiti

Davies

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

103

120

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“…A recent report suggests that UCHL1 can inhibit MT formation in a ubiquitination-dependent manner (Bheda et al, 2010). Up-regulation of this enzyme may partly contribute to the depolymerization of MTs observed in our study.…”

Section: Discussionmentioning

confidence: 44%

2D-DIGE Proteomic Analysis of Mesenchymal Stem Cell Cultured on the Elasticity-tunable Hydrogels

Kuboki

Kantawong

Burchmore

et al. 2012

Cell Struct. Funct.

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ABSTRACT. The present study focuses on mechanotransduction in mesenchymal stem cells (MSCs) in response to matrix elasticity. By using photocurable gelatinous gels with tunable stiffness, proteomic profiles of MSCs cultured on tissue culture plastic, soft (3 kPa) and stiff (52 kPa) matrices were deciphered using 2-dimensional differential in-gel analysis (2D-DIGE). The DIGE data, tied to immunofluorescence, indicated abundance and organization changes in the cytoskeletonal proteins as well as differential regulation of important signalingrelated proteins, stress-responsing proteins and also proteins involved in collagen synthesis. The major CSK proteins including actin, tubulin and vimentin of the cells cultured on the gels were remarkably changed their expressions. Significant down-regulation of α-tubulin and β-actin can be observed on gel samples in comparison to the rigid tissue culture plates. The expression abundance of vimentin appeared to be highest in the MSCs cultured on hard gels. These results suggested that the substrate stiffness significantly affects expression balances in cytoskeletal proteins of MSCs with some implications to cellular tensegrity.

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“…The colocalization of PTOV1 and UCH-L1 in oocyte nuclei may be involved in regulating meiotic division. UCH-L1 is tightly associated with the mitotic spindle through all stages of M phase, suggesting that it plays an important role in microtubule formation (Bheda et al 2010). Nuclear localization of PTOV1 is required for the stimulation of cell proliferation (Santamaria et al 2005;Santamaria et al 2003).…”

Section: Discussionmentioning

confidence: 99%

PTOV1 is associated with UCH-L1 and in response to estrogen stimuli during the mouse oocyte development

Shi

Jia

et al. 2011

Histochem Cell Biol

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To investigate the biological significance of ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) involvement in oocyte maturation, we screened for proteins that bound to UCH-L1 in mouse ovaries, and we found that the prostate tumor overexpressed-1 (PTOV1) protein was able to bind to UCH-L1. PTOV1 is highly expressed in prostate cancers and considered as a potential marker for carcinogenesis and the progress of prostate cancer. It was reported that PTOV1 plays an important role in cell cycle regulation, but its role in mammalian oocyte development and meiosis is still unclear. In this paper, it was found that the expression levels of PTOV1 in mouse ovaries progressively increased from prepubescence to adulthood. And we found by immunohistochemistry that PTOV1 spreaded in both the cytoplasm and nuclei of oocytes during prepuberty, but in normal adult mouse oocytes, it concentrated not only in nuclei but also on the plasma membrane, though in some oocytes with abnormal shapes, PTOV1 did not display the typical distribution patterns. In granulosa cells, however, it was found to locate in the cytoplasm at all the selected ages. In postnatal mouse ovaries (28 days), estradiol treatment induced the adult-specific distribution pattern of PTOV1 in oocytes. In addition, UCH-L1 was shown to be associated with CDK1, which participated in the regulation of cell cycle and oocyte maturation. Therefore, we propose that the distribution changes of PTOV1 are age-dependent, and significant for mouse oocyte development and maturation. Moreover, the discovery that PTOV1 is associated with UCH-L1 in mouse oocytes supports the explanations for that UCH-L1 is involved in oocyte development and maturation, especially under the regulation of estrogen.

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Ubiquitin editing enzyme UCH L1 and microtubule dynamics: Implication in mitosis

Cited by 60 publications

References 46 publications

Ubiquitin vinyl methyl ester binding orients the misaligned active site of the ubiquitin hydrolase UCHL1 into productive conformation

Ubiquitin vinyl methyl ester binding orients the misaligned active site of the ubiquitin hydrolase UCHL1 into productive conformation

2D-DIGE Proteomic Analysis of Mesenchymal Stem Cell Cultured on the Elasticity-tunable Hydrogels

PTOV1 is associated with UCH-L1 and in response to estrogen stimuli during the mouse oocyte development

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