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J Biol Chem ; 279(21): 22362-70, 2004 May 21.
Artículo en Inglés | MEDLINE | ID: mdl-14996835

RESUMEN

dUTPase prevents uracil incorporation into DNA by strict regulation of the cellular dUTP:dTTP ratio. Lack of the enzyme initiates thymineless cell death, prompting studies on enzyme regulation. We investigated expression pattern and localization of Drosophila dUTPase. Similarly to human, two isoforms of the fly enzyme were identified at both mRNA and protein levels. During larval stages, a drastic decrease of dUTPase expression was demonstrated at the protein level. In contrast, dUTPase mRNAs display constitutive character throughout development. A putative nuclear localization signal was identified in one of the two isoforms. However, immunohistochemistry of ovaries and embryos did not show a clear correlation between the presence of this signal and subcellular localization of the protein, suggesting that the latter may be perturbed by additional factors. Results are in agreement with a multilevel regulation of dUTPase in the Drosophila proteome, possibly involving several interacting protein partners of the enzyme. Using independent approaches, the existence of such macromolecular partners was verified.


Asunto(s)
Drosophila melanogaster/enzimología , Pirofosfatasas/biosíntesis , Pirofosfatasas/química , Secuencia de Aminoácidos , Animales , Secuencia de Bases , Western Blotting , Ciclo Celular , Muerte Celular , Línea Celular , Drosophila , Femenino , Regulación del Desarrollo de la Expresión Génica , Inmunohistoquímica , Hibridación in Situ , Espectrometría de Masas , Microscopía Fluorescente , Datos de Secuencia Molecular , Hibridación de Ácido Nucleico , Ovario/metabolismo , Pruebas de Precipitina , Isoformas de Proteínas , Proteoma , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Fracciones Subcelulares/metabolismo , Resonancia por Plasmón de Superficie , Factores de Tiempo
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