Expression and Functional Analysis of Uch-L3 during Mouse Development

Howard Hughes Medical Institute and Department of Molecular Biology, Princeton University, Princeton, New Jersey 08544, USA.
Molecular and Cellular Biology (Impact Factor: 4.78). 05/2000; 20(7):2498-504. DOI: 10.1128/MCB.20.7.2498-2504.2000
Source: PubMed

ABSTRACT Mice homozygous for the s(1Acrg) deletion at the Ednrb locus arrest at embryonic day 8.5. To determine the molecular basis of this defect, we initiated positional cloning of the s(1Acrg) minimal region. The mouse Uch-L3 (ubiquitin C-terminal hydrolase L3) gene was mapped within the s(1Acrg) minimal region. Because Uch-L3 transcripts were present in embryonic structures relevant to the s(1Acrg) phenotype, we created a targeted mutation in Uch-L3 to address its role during development and its possible contribution to the s(1Acrg) phenotype. Mice homozygous for the mutation Uch-L3(Delta3-7) were viable, with no obvious developmental or histological abnormalities. Although high levels of Uch-L3 RNA were detected in testes and thymus, Uch-L3(Delta3-7) homozygotes were fertile, and no defect in intrathymic T-cell differentiation was detected. We conclude that the s(1Acrg) phenotype is either complex and multigenic or due to the loss of another gene within the region. We propose that Uch-L3 may be functionally redundant with its homologue Uch-L1.

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Available from: Ekaterina Semenova, Sep 26, 2015
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    • "A unique feature of UCH-L3 is that this enzyme has dual hydrolase specificity towards Ub and Nedd8 [8]. UCH-L3 is expressed ubiquitously in all tissues but the expression of UCH-L1 is restricted to the testes, ovaries, and neurons [4] [6] [9]. "
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    ABSTRACT: Ubiquitin C-terminal hydrolse-L3 (UCH-L3) is among the deubiquitinating enzymes (DUBs) that cleave ubiquitin (Ub) from Ub precursors or protein substrates. Many DUBs have been shown to participate in cancer progression in various tissues. However, the mechanism and role of UCH-L3 in carcinogenesis has largely been unknown until recently. Here we investigated the implication of UCH-L3 in prostate cancer progression. Interestingly, UCH-L3 is upregulated in normal or non-metastatic prostate cancer cells and is downregulated in metastatic prostate cancer cell lines. Notably, knockdown of UCH-L3 in normal prostate cell line RWPE1 promotes epithelial-to-mesenchymal transition (EMT), an important process for cancer cell invasion and metastasis. The induction of EMT by UCH-L3 knockdown results in an increase of cell migration and invasion. Yet, to the contrary, overexpression of UCH-L3 in highly metastatic prostate cancer cell line PC3 reverses EMT but the active site mutant UCH-L3 didn't. Collectively, our findings identify UCH-L3 as a novel EMT regulator in prostate cells and highlight UCH-L3 as a potential therapeutic target for preventing metastatic prostate cancer.
    Biochemical and Biophysical Research Communications 09/2014; 452(3). DOI:10.1016/j.bbrc.2014.08.144 · 2.30 Impact Factor
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    • "Meiotic pachytene spermatocytes and postmeiotic spermatids; cauda epididymis Sperm quality control during epididymal maturation Mus musculus [48] [109] [110] Uch-L4 All tissues, with testis included Mus musculus [111] Uch-L5 Spermatocytes and spermatids Mus musculus [112] CYLD Control of spermatogenetic cell apoptosis and spermatogenesis progression via RIP1/NF-kappaB signalling axis Mus musculus [113] * Indicates " also known as DnaJB13 " . "
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    ABSTRACT: Spermatogenesis is a complex process in which mitosis, meiosis, and cell differentiation events coexist. The need to guarantee the production of qualitatively functional spermatozoa has evolved into several control systems that check spermatogenesis progression/sperm maturation and tag aberrant gametes for degradation. In this review, we will focus on the importance of the evolutionarily conserved molecular pathways involving molecular chaperones belonging to the superfamily of heat shock proteins (HSPs), their cochaperones, and ubiquitination/deubiquitination system all over the spermatogenetic process. In this respect, we will discuss the conserved role played by the DNAJ protein Msj-1 (mouse sperm cell-specific DNAJ first homologue) and the deubiquitinating enzyme Ubpy (ubiquitin-specific processing protease-y) during the spermiogenesis in both mammals and nonmammalian vertebrates.
    BioMed Research International 06/2014; 2014(10):561426. DOI:10.1155/2014/561426 · 2.71 Impact Factor
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    • "To date, the only isopeptidase known to function exclusively in NEDD8 processing and deconjugation is the C48 family peptidase DEN1/NEDP1/SENP8 from Drosophila, and human (Figure 3; Gan-Erdene et al., 2003; Mendoza et al., 2003; Wu et al., 2003; Shen et al., 2005; Chan et al., 2008; Shin et al., 2011). However, mouse knockouts of UCHL3 or Drosophila and Aspergillus knockouts of DEN1 are viable although NEDD8 and neddylation are essential in the respective organisms (Kurihara et al., 2000; Chan et al., 2008; Christmann et al., 2013). These findings suggest that mutants of these processing enzymes cannot be fully impaired in NEDD8 processing. "
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    ABSTRACT: NEDD8, in plants and yeasts also known as RELATED TO UBIQUITIN (RUB), is an evolutionarily conserved 76 amino acid protein highly related to ubiquitin. Like ubiquitin, NEDD8 can be conjugated to and deconjugated from target proteins, but unlike ubiquitin, NEDD8 has not been reported to form chains similar to the different polymeric ubiquitin chains that have a role in a diverse set of cellular processes. NEDD8-modification is best known as a post-translational modification of the cullin subunits of cullin-RING E3 ubiquitin ligases. In this context, structural analyses have revealed that neddylation induces a conformation change of the cullin that brings the ubiquitylation substrates into proximity of the interacting E2 conjugating enzyme. In turn, NEDD8 deconjugation destabilizes the cullin RING ligase complex allowing for the exchange of substrate recognition subunits via the exchange factor CAND1. In plants, components of the neddylation and deneddylation pathway were identified based on mutants with defects in auxin and light responses and the characterization of these mutants has been instrumental for the elucidation of the neddylation pathway. More recently, there has been evidence from animal and plant systems that NEDD8 conjugation may also regulate the behavior or fate of non-cullin substrates in a number of ways. Here, the current knowledge on NEDD8 processing, conjugation and deconjugation is presented, where applicable, in the context of specific signaling pathways from plants.
    Frontiers in Plant Science 03/2014; 5:103. DOI:10.3389/fpls.2014.00103 · 3.95 Impact Factor
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