title: Drosophila nuclear factor DREF regulates the expression of the mitochondrial DNA helicase and mitochondrial transcription factor B2 but not the mitochondrial translation factor B1.
abstract: DREF [DRE (DNA replication-related element)-binding factor] controls the transcription of numerous genes in Drosophila, many involved in nuclear DNA (nDNA) replication and cell proliferation, three in mitochondrial DNA (mtDNA) replication and two in mtDNA transcription termination. In this work, we have analysed the involvement of DREF in the expression of the known remaining genes engaged in the minimal mtDNA replication (d-mtDNA helicase) and transcription (the activator d-mtTFB2) machineries and of a gene involved in mitochondrial mRNA translation (d-mtTFB1). We have identified their transcriptional initiation sites and DRE sequences in their promoter regions. Gel-shift and chromatin immunoprecipitation assays demonstrate that DREF interacts in vitro and in vivo with the d-mtDNA helicase and d-mtTFB2, but not with the d-mtTFB1 promoters. Transient transfection assays in Drosophila S2 cells with mutated DRE motifs and truncated promoter regions show that DREF controls the transcription of d-mtDNA helicase and d-mtTFB2, but not that of d-mtTFB1. RNA interference of DREF in S2 cells reinforces these results showing a decrease in the mRNA levels of d-mtDNA helicase and d-mtTFB2 and no changes in those of the d-mtTFB1. These results link the genetic regulation of nuclear DNA replication with the genetic control of mtDNA replication and transcriptional activation in Drosophila.
authors.affiliation: Departamento de Bioquímica, Instituto de Investigaciones Biomédicas "Alberto Sols" UAM-CSIC and Centro de Investigación Biomédica en Red (CIBERER), Facultad de Medicina, Universidad Autónoma de Madrid, Spain, c/ Arzobispo Morcillo 4, 28029 Madrid, Spain; Instituto de Investigación Sanitaria Hospital Universitario 12 de Octubre (i+12), Madrid, Spain. Electronic address: miguel.fernandez@uam.es.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Fernández-Moreno
authors.forename: Rosana
authors.initials: R
authors.lastname: Hernández
authors.forename: Cristina
authors.initials: C
authors.lastname: Adán
authors.forename: Marina
authors.initials: M
authors.lastname: Roberti
authors.forename: Francesco
authors.initials: F
authors.lastname: Bruni
authors.forename: Paola Loguercio
authors.initials: PL
authors.lastname: Polosa
authors.forename: Palmiro
authors.initials: P
authors.lastname: Cantatore
authors.forename: Yuichi
authors.initials: Y
authors.lastname: Matsushima
authors.forename: Laurie S
authors.initials: LS
authors.lastname: Kaguni
authors.forename: Rafael
authors.initials: R
authors.lastname: Garesse
mesh_terms: D000818:Animals; D015153:Blotting, Western; D002467:Cell Nucleus; D047369:Chromatin Immunoprecipitation; D004265:DNA Helicases / Q000235:genetics* / Q000378:metabolism; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D004331:Drosophila melanogaster / Q000235:genetics*; D024202:Electrophoretic Mobility Shift Assay; D005786:Gene Expression Regulation; D008156:Luciferases; D008928:Mitochondria / Q000235:genetics / Q000378:metabolism*; D024101:Mitochondrial Proteins / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism; D024363:Transcription Initiation Site
chemical_list: C099494:Dref protein, Drosophila; D029721:Drosophila Proteins; D024101:Mitochondrial Proteins; D012333:RNA, Messenger; D014157:Transcription Factors; D008156:Luciferases; D004265:DNA Helicases
keywords: DRE (DNA replication-related element)-binding factor; DREF; Drosophila; Drosophila mitochondrial transcription factor B1; Drosophila mitochondrial transcription factor B2; Mitochondrial biogenesis; Nuclear DNA replication; Transcriptional regulation; d-mtTFB1; d-mtTFB2; mitochondrial mRNAs; mtmRNA; nts; nucleotides; transcription start site; tss
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