RESUMO

Dystrophin Dp40 is the shortest protein encoded by the DMD (Duchenne muscular dystrophy) gene. This protein is unique since it lacks the C-terminal end of dystrophins. In this data article, we describe the subcellular localization, nuclear export signals and the three-dimensional structure modeling of putative Dp40 proteins using bioinformatics tools. The Dp40 wild type protein was predicted as a cytoplasmic protein while the Dp40n4 was predicted to be nuclear. Changes L93P and L170P are involved in the nuclear localization of Dp40n4 protein. A close analysis of Dp40 protein scored that amino acids (93)LEQEHNNLV(101) and (168)LLLHDSIQI(176) could function as NES sequences and the scores are lost in Dp40n4. In addition, the changes L93/170P modify the tertiary structure of putative Dp40 mutants. The analysis showed that changes of residues 93 and 170 from leucine to proline allow the nuclear localization of Dp40 proteins. The data described here are related to the research article entitled "EF-hand domains are involved in the differential cellular distribution of dystrophin Dp40" (J. Aragón et al. Neurosci. Lett. 600 (2015) 115-120) [1].

RESUMO

Dp40 is the shortest DMD gene product that has been reported to date. It is encoded by exons 63-70, a region required for a ß-dystroglycan interaction. Its expression has been identified in rat, mouse, and human; however, its function remains unknown. To explore the expression of Dp40 transcript and subcellular localization of epitope-tagged Dp40 proteins, RT-PCR and immunofluorescence assays were performed in PC12 cells. The expression of Dp40 mRNA was found in undifferentiated and nerve growth factor-differentiated PC12 cells. According to immunofluorescence analyses, the recombinant protein Dp40 was mainly localized in the cell periphery/cytoplasm of undifferentiated and differentiated PC12 cells, a small amount of this protein is localized to the nucleus of differentiated cells. With the aim to identify the amino acids involved in the nuclear localization of Dp40, an in silico analysis was performed and it predicted that prolines 93 and 170, located within EF1 and EF2-hand domains, are involved in the nuclear localization of this protein. This prediction was confirmed by site-directed mutagenesis, the Dp40-L93P mutant was localized to the nucleus and cell periphery, while Dp40-L170P and Dp40-L93/170P showed mainly a nuclear localization. Dp40 co-localizes with ß-dystroglycan and the co-localization score was statistically reduced in Dp40-L93P, Dp40-L170P and Dp40-L93/170P mutants.

Assuntos

Distrofina/metabolismo , Animais , Diferenciação Celular , Núcleo Celular/metabolismo , Distroglicanas/metabolismo , Distrofina/genética , Células HeLa , Humanos , Mutagênese Sítio-Dirigida , Mutação , Células PC12 , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Estrutura Terciária de Proteína , Ratos

RESUMO

Hace referencia a varias alternativas de ampliacion de plataforma, indicando el lado favorable y desfavorable de cada uno de ellos por ultimo se hace un enfoque de la alternativa propuesta.El objetivo del proyecto propuesto es evitar grandes movimientos de tierra que desestabilizan la estructura, se halla ubicado en la carretera de Cotapata-Santa Barbara Km 38+840. El desarrollo del proyecto se realizo basandose en las recomendaciones de la AASHTO y otros textos para el diseno de estructuras de hormigon armado. Finalmente se realizan los presupuestos del proyecto inicial y alternativo por lo que se llega a la conclusion de que el proyecto alternativo es favorable tanto en el aspecto tecnico como economico.