RESUMO
The molecular etiology of obesity predisposition is largely unknown. Here, we present evidence that genetic variation in TBC1D1 confers risk for severe obesity in females. We identified a coding variant (R125W) in TBC1D1 that segregated with the disease in 4p15-14-linked obesity pedigrees. In cases derived from pedigrees with the strongest linkage evidence, the variant was significantly associated with obesity (P=0.000007) and chromosomes carrying R125W accounted for the majority of the evidence that originally linked 4p15-14 with the disease. In addition, by selecting families that segregated R125W with obesity, we were able to generate highly significant linkage evidence for an obesity predisposition locus at 4q34-35. This result provides additional and confirming evidence that R125W affects obesity susceptibility, delimits the location of an obesity gene at 4q34-35 and identifies a gene/gene interaction that influences the risk for obesity predisposition. Finally, although the function of TBC1D1 is unknown, the protein is structurally similar to a known regulator of insulin-mediated Glut4 translocation.
Assuntos
Endopeptidases/genética , Obesidade/genética , Proteínas Oncogênicas/genética , Cromossomos Humanos Par 4/genética , Feminino , Expressão Gênica , Variação Genética , Haplótipos , Humanos , Desequilíbrio de Ligação , Escore Lod , Masculino , Obesidade/etiologia , Linhagem , Fenótipo , Polimorfismo de Nucleotídeo Único , Proteínas Proto-Oncogênicas , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Tecidual , Ubiquitina TiolesteraseRESUMO
HIV release requires TSG101, a cellular factor that sorts proteins into vesicles that bud into multivesicular bodies (MVB). To test whether other proteins involved in MVB biogenesis (the class E proteins) also participate in HIV release, we identified 22 candidate human class E proteins. These proteins were connected into a coherent network by 43 different protein-protein interactions, with AIP1 playing a key role in linking complexes that act early (TSG101/ESCRT-I) and late (CHMP4/ESCRT-III) in the pathway. AIP1 also binds the HIV-1 p6(Gag) and EIAV p9(Gag) proteins, indicating that it can function directly in virus budding. Human class E proteins were found in HIV-1 particles, and dominant-negative mutants of late-acting human class E proteins arrested HIV-1 budding through plasmal and endosomal membranes. These studies define a protein network required for human MVB biogenesis and indicate that the entire network participates in the release of HIV and probably many other viruses.
Assuntos
Membrana Celular/virologia , HIV-1/metabolismo , Proteínas/metabolismo , Vesículas Transportadoras/virologia , Eliminação de Partículas Virais/fisiologia , Animais , Células COS , Compartimento Celular/genética , Linhagem Celular , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Proteínas de Ligação a DNA/metabolismo , Complexos Endossomais de Distribuição Requeridos para Transporte , Endossomos/genética , Endossomos/metabolismo , Endossomos/ultraestrutura , Produtos do Gene gag/metabolismo , HIV-1/genética , HIV-1/ultraestrutura , Humanos , Proteínas dos Microfilamentos/metabolismo , Microscopia Eletrônica , Modelos Biológicos , Mutação/genética , Ligação Proteica/fisiologia , Fatores de Transcrição/metabolismo , Vesículas Transportadoras/metabolismo , Vesículas Transportadoras/ultraestrutura , Proteínas Virais/metabolismo , Produtos do Gene gag do Vírus da Imunodeficiência HumanaRESUMO
The HIV-1 Gag protein recruits the cellular factor Tsg101 to facilitate the final stages of virus budding. A conserved P(S/T)AP tetrapeptide motif within Gag (the "late domain") binds directly to the NH2-terminal ubiquitin E2 variant (UEV) domain of Tsg101. In the cell, Tsg101 is required for biogenesis of vesicles that bud into the lumen of late endosomal compartments called multivesicular bodies (MVBs). However, the mechanism by which Tsg101 is recruited from the cytoplasm onto the endosomal membrane has not been known. Now, we report that Tsg101 binds the COOH-terminal region of the endosomal protein hepatocyte growth factor-regulated tyrosine kinase substrate (Hrs; residues 222-777). This interaction is mediated, in part, by binding of the Tsg101 UEV domain to the Hrs 348PSAP351 motif. Importantly, Hrs222-777 can recruit Tsg101 and rescue the budding of virus-like Gag particles that are missing native late domains. These observations indicate that Hrs normally functions to recruit Tsg101 to the endosomal membrane. HIV-1 Gag apparently mimics this Hrs activity, and thereby usurps Tsg101 and other components of the MVB vesicle fission machinery to facilitate viral budding.