RESUMO
The human ubiquitin-homology domain protein PIC1 interacts with the acute promyelocytic leukemia protein PML, and both proteins form part of the large, nuclear, multiprotein complexes known as PML nuclear bodies. The normal punctate immunohistochemical staining pattern of these complexes is disrupted by viral infection or interferon treatment and in blast cells from patients with acute promyelocytic leukemia. We have characterized the murine homologue of PIC1 and have found that the predicted amino acid sequences of the mouse and human proteins are identical. High levels of Pic1 mRNA were detected in a range of mouse tissues. Pic1 genomic clones were isolated, and the organization of the gene was determined. Two processed Pic1 pseudogenes were also isolated and characterized. Through FISH, the chromosomal localizations of the mouse Pic1 gene and the two pseudogenes were determined. Human PIC1 (HGMW-approved symbol UBL1)-related sequences were isolated from human genomic DNA and were shown to represent processed pseudogenes. The role of PIC1 in a variety of cellular processes is discussed.
Assuntos
Proteínas de Transporte/genética , Pseudogenes , Ubiquitinas/genética , Células 3T3/imunologia , Células 3T3/metabolismo , Sequência de Aminoácidos , Animais , Mapeamento Cromossômico , Sequência Conservada , Reações Cruzadas , Éxons , Humanos , Hibridização In Situ/métodos , Íntrons , Camundongos , Dados de Sequência Molecular , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/metabolismo , Proteína SUMO-1 , Homologia de Sequência de Aminoácidos , Ubiquitinas/imunologiaRESUMO
Organotypic cultures of human skin were made using dermal fibroblasts seeded into a type I collagen gel overlaid with epidermal keratinocytes. Full-thickness excision of tattoos was performed on five patients, three of whom received sex-mismatched allografts. Patients were not immunosuppressed. Biopsies were obtained up to 3.5 years later. In situ hybridization of the PHY2.1 repetitive Y chromosome sequence revealed male fibroblasts and keratinocytes at 11 weeks and 2.5 years in the two female patients grafted with male cells. Structural components in the dermal substitute matured with time, and elastic fibers formed an interlacing meshwork by 18 months. Electron microscopy of the dermal-epidermal junction of an organotypic allograft revealed anchoring fibrils that had normal features at this time. Hyperemia of early grafts settled and contour correction was maintained, while repigmentation was variable. Hypertrophic scars did not occur, and graft contracture was never more than 20 percent. We conclude that this organotypic skin graft shows potential toward the goal of allogeneic skin replacement in a one-step procedure.
