Molecular characterization and developmental expression of the TFIIH factor p62 gene from Drosophila melanogaster: effects on the UV light sensitivity of a p62 mutant fly.
TFIIH is a multiprotein complex that has a central role in the RNA pol II mediated transcription, in DNA repair and in the control of the cell cycle. Mutations in some components of TFIIH are associated with three hereditary human syndromes: xeroderma pigmentosum (XP), Cockayne syndrome (CS) and trichothiodystrophy (TTD). The p62 protein is a structural component of the TFIIH core and no syndromes have been identified up to date by mutations in this human gene. In this work we report the molecular and genetic characterization of the Drosophila melanogaster p62 gene (Dmp62). The Dmp62 gene product shows high identity with its human and mouse homologues. Using computer analysis we identified several common motifs in the p62 proteins from different organisms, suggesting that these motifs could be involved in possible protein-protein interactions within the TFIIH complex or with other transcription and DNA repair factors. The Dmp62 transcript is expressed at similar levels throughout development, although there is a significant increase of the transcript level during the late embryogenesis and in the adult male. The analysis of a Drosophila line with a P-element enhancer trap insertion at the Dmp62 5'-UTR that directs the lac-Z expression from the Dmp62 promoter, showed a high level of expression in the gut, the testis and the pericardial cells. A P-element that disrupts the Dmp62 gene (Dmp62mut) produces early embryo lethality in homozygous flies. Heterozygous Dmp62mut larvae are more sensitive to UV light irradiation, and those individuals that are able to develop into adults have severe abdominal cuticular damage after UV light irradiation.
Castro
Juan
J
Department of Genetics and Molecular Physiology, Institute of Biotechnology, Universidad Nacional Autónoma de México, Cuernavaca, Morelos 62250, Mexico.
Merino
Carlos
C
Zurita
Mario
M
D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017931:DNA Primers / Q000737:chemistry; D004260:DNA Repair / Q000235:genetics; D029721:Drosophila Proteins / Q000096:biosynthesis / Q000235:genetics*; D004331:Drosophila melanogaster / Q000235:genetics / Q000528:radiation effects*; D004625:Embryo, Nonmammalian / Q000378:metabolism; D004742:Enhancer Elements, Genetic; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D007763:Lac Operon; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D035381:TATA-Binding Protein Associated Factors; D014018:Tissue Distribution; D035362:Transcription Factor TFIID; D051758:Transcription Factor TFIIH; D019473:Transcription Factors, TFII / Q000096:biosynthesis / Q000235:genetics*; D014466:Ultraviolet Rays; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D029721:Drosophila Proteins; D012333:RNA, Messenger; D035381:TATA-Binding Protein Associated Factors; C084976:Taf6 protein, Drosophila; D035362:Transcription Factor TFIID; D019473:Transcription Factors, TFII; D051758:Transcription Factor TFIIH; D001616:beta-Galactosidase