Nuclear domain 10 (ND10), also referred to as nuclear bodies, are discrete interchromosomal accumulations of several proteins including promyelocytic leukemia protein (PML) and Sp100. In this study, we investigated the mechanism ND10 assembly by identifying that essential for process using cells lines lack individual ND10-associated proteins. We identified adapter Daxx BML, RecQ helicase missing in Bloom syndrome, new PML, but not BLM or Sp100, was found be responsible proper localization...

Fas/APO-1 and TNF receptor 1 share a common signaling motif in their cytoplasmic tail called the "death domain." Using death domain as bait yeast two-hybrid system, several domain-containing proteins that participate cell have been identified. Here we report isolation of novel protein, sentrin, which interacts with but not FADD/MORT1 or CD40. Two-hybrid interaction assays reveal sentrin associates only signal-competent forms domains. Sentrin is protein 101 amino acids homology to ubiquitin,...

Acute promyelocytic leukemia arises following a reciprocal chromosome translocation t(15;17), which generates PML-retinoic acid receptor alpha fusion proteins (PML-RARalpha). We have shown previously that wild type PML, but not PML-RARalpha, is covalently modified by the sentrin family of ubiquitin-like (Kamitani, T., Nguyen, H. P., Kito, K., Fukuda-Kamitani, and Yeh, E. T. (1998) J. Biol. Chem. 273, 3117-3120). To understand mechanisms underlying differential sentrinization PML versus...

PML, a RING finger protein with tumor suppressor activity, has been implicated in the pathogenesis of acute promyelocytic leukemia that arises following reciprocal chromosomal translocation fuses PML gene retinoic acid receptor α (RARα) gene. Immunocytochemical analysis demonstrated is co-localized novel ubiquitin-like nuclear bodies, which could be disrupted by PML-RARα fusion protein. The physical nature this co-localization unknown. Using COS cell expression system, we show covalently...

Sentrin is a ubiquitin-like molecule that has been shown to interact with the death domains of Fas and tumor necrosis factor receptor 1 (TNFR1), PML, Rad51, Rad52, RanGAP1. We have reported previously sentrin can be conjugated other proteins in manner analogous protein ubiquitination (Kamitani, T., Nguyen, H. P., Yeh, E. T. (1997) J. Biol. Chem. 272, 14001-14004). Furthermore, conserved C-terminal Gly-Gly residues are required for sentrinization occur. To identify enzymes which play role...

α-Synuclein is a key molecule in understanding the pathogenesis of neurodegenerative α-synucleinopathies such as Parkinson's disease. Despite extensive research, however, its precise function remains unclear partly because difficulty immunoblotting detection endogenous α-synuclein. This has largely restricted progress for α-synucleinopathy research. Here, we report that α-synuclein monomers tend to easily detach from blotted membranes, resulting no or very poor detection. To prevent this...

Sentrin is a novel ubiquitin-like protein that protects cells against both anti-Fas and tumor necrosis factor-induced cell death. Antiserum recognizing the N terminus of sentrin revealed presence 18-kDa monomer, 90-kDa band (p90), multiple high molecular mass bands. Because possesses conserved Gly-Gly residues near C terminus, it likely these additional bands represent conjugation to other proteins in manner similar ubiquitination pathway. Transient expression hemagglutinin epitope-tagged...

NEDD8 is a ubiquitin-like protein that controls vital biological events through its conjugation to cullin family members. Recently, we identified negative regulator of the system, NUB1, which interacts with and down-regulates expression post-transcriptionally (Kito, K., Yeh, E. T. H., Kamitani, (2001) <i>J. Biol. Chem.</i> 276, 20603–20609). Here, show NUB1 possesses domain at N-terminal region binds S5a 19 S proteasome activator (PA700). A GST pull-down assay revealed overexpression leads...

Sentrin is a novel ubiquitin-like protein that can be conjugated to other proteins in manner analogous ubiquitination. Two additional cDNA sequences encode highly homologous sentrin have been reported GenBank™. It not known whether these sentrin-like could also function as modifiers. In this report, second member of the family was characterized detail. Sentrin-2 95-amino acid polypeptide 46% identical and 66% sentrin-1. Northern blot analysis showed sentrin-2 message expressed all tissues,...

HIV‐1 efficiently infects susceptible cells and causes AIDS in humans. Although HIV can also enter the of Old World monkeys, it encounters a block before reverse transcription. Data have shown that this species‐specific restriction is mediated by tripartite motif (TRIM)5α, whose molecular function still undefined. Here, we show TRIM5α functions as RING‐finger‐type E3 ubiquitin ligase both vitro vivo ubiquitinates itself cooperation with E2 ubiquitin‐conjugating enzyme UbcH5B. In addition to...

Melanoma is the major cause of skin cancer death worldwide. Parkinson's disease a neurodegenerative disorder that caused by mutation alpha-synuclein or other genes. Importantly, epidemiological studies have reported co-occurrence melanoma and disease, suggesting these two diseases could share common genetic components.Recently, we found human cell lines highly express alpha-synuclein, whereas protein undetectable in non-melanoma tested. To investigate expression tissues, immunostained...

NEDD8 (neural precursor cell expressed, developmentally down‐regulated 8) is a ubiquitin‐like protein that controls vital biological events through its conjugation to members of the cullin family, which are components certain ubiquitin E3 ligases. Recent studies have shown incorporated into Lewy bodies (LBs) in Parkinson's disease, Mallory alcoholic liver disease and Rosenthal fibres astrocytoma. In order examine whether plays role formation ubiquitinated inclusions, we performed...

F'revious attempts to express glycosylphosphatidylinositol-anchored proteins in Ltk-cells have not been successful because cannot synthesizeN-acetylglucosamine-phosphatidylinositol, the first intermediate anchor biosynthesis.Using complementation cloning, we identified a human cDNA that corrects defect biosynthesis and allows expression of Ltk-cells.The nucleotide sequence predicts novel cytosolic protein 188 amino acids.Glycosylphosphatidylinositol (GPI)' serves as membrane for large number...

ABSTRACT Nitric oxide (NO · ) produced by inducible nitric synthase (iNOS) is an important host defense molecule against Mycobacterium tuberculosis in mononuclear phagocytes. The objective of this study was to determine the role IκBα kinase-nuclear factor κB (IKK-NF-κB) signaling pathway induction iNOS and NO a mycobacterial cell wall lipoglycan known as mannose-capped lipoarabinomannan (ManLAM) mouse macrophages costimulated with gamma interferon (IFN-γ). NF-κB activated ManLAM shown...

Fas (APO1/CD95) is a type 1 transmembrane protein critically involved in receptor-mediated apoptosis. Previous studies have shown that exists monomeric form resting cells and aggregates upon cross-linking to complex serves recruit additional signaling molecules the cell membrane. To study molecular fate of antigen following receptor activation, monoclonal antibody specific for death domain has been generated. This (3D5) could be used Western blot analysis using total lysates identify...