Unknown,Transcriptomics,Genomics,Proteomics

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Functional characterization of a Drosophila melanogaster mitochondrial deoxynucleotide transporter dRIM2.

ABSTRACT: We have studied the deoxynucleotide transport in Drosophila melanogaster. On the basis of homology with the S. cerevisiae RIM2 gene, encoding a pyrimidine deoxynucleotide carrier (Marrobbio et al. 2006), the CG18317 gene (dRIM2) in the fruit fly may code for a deoxynucleotide carrier. We demonstrated that Drosophila S2R+ cells, silenced for the dRIM2 expression, had a marked defect in the amounts of all mitochondrial dNTPs, both purines and pyrimidines. In vivo dRIM2 homozygous knockout produced a larval lethal phenotype. dRIM2-/- larvae showed (i) impairments in the locomotor behavior, (ii) a decrease in the rates of oxygen consumption and (iii) a depletion of the mtDNA. Following a detailed morphological characterization carried out in dRIM2-/- larvae evidenced an ongoing mitochondrial biogenesis accompanied by an alteration of mitochondria shaping. Additionally, the role of dRIM2 in the purine and pyrimidine metabolism was supported by a microarray analysis. We conclude that dRIM2 is a Drosophila deoxynucleotide carrier, essential for maintaining the mitochondrial functionality. Gene expression profiling was carried out on dRIM-/- and dRIM+/- Drosophila larvae using the Drosophila 1.0 custom platform (Agilent). Total RNA was obtained from the whole body of 3rd instar larvae for each genotype. Four and three biological replicates were analyzed for dRIM-/- and dRIM+/- samples respectively for a total of 7 microarray experiments.

ORGANISM(S): Drosophila melanogaster

SUBMITTER: Cristiano De Pitta

PROVIDER: E-GEOD-48012 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

Functional characterization of drim2, the Drosophila melanogaster homolog of the yeast mitochondrial deoxynucleotide transporter.

Da-Rè Caterina C Franzolin Elisa E Biscontin Alberto A Piazzesi Antonia A Pacchioni Beniamina B Gagliani Maria Cristina MC Mazzotta Gabriella G Tacchetti Carlo C Zordan Mauro A MA Zeviani Massimo M Bernardi Paolo P Bianchi Vera V De Pittà Cristiano C Costa Rodolfo R

The Journal of biological chemistry 20140127 11

The CG18317 gene (drim2) is the Drosophila melanogaster homolog of the Saccharomyces cerevisiae Rim2 gene, which encodes a pyrimidine (deoxy)nucleotide carrier. Here, we tested if the drim2 gene also encodes for a deoxynucleotide transporter in the fruit fly. The protein was localized to mitochondria. Drosophila S2R(+) cells, silenced for drim2 expression, contained markedly reduced pools of both purine and pyrimidine dNTPs in mitochondria, whereas cytosolic pools were unaffected. In vivo drim2 ...[more]

PMID: 24469456

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Project description:BACKGROUND: Polar environments are characterized by extreme seasonal changes in day length, light intensity and spectrum, the extent of sea ice during the winter, and food availability. A key species of the Southern Ocean ecosystem, the Antarctic krill (Euphausia superba) has evolved rhythmic physiological and behavioral mechanisms to adapt to daily and seasonal changes. The molecular organization of the clockwork underlying these biological rhythms is, nevertheless, still only partially understood. METHODOLOGY/PRINCIPAL FINDINGS:The genome sequence of the Antarctic krill is not yet available. A normalized cDNA library was produced and pyrosequenced in the attempt to identify large numbers of transcripts. All available E. superba sequences were then assembled to create the most complete existing oligonucleotide microarray platform with a total of 32,217 probes. Gene expression signatures of specimens collected in the Ross Sea at five different time points over a 24-hour cycle were defined, and 1,308 genes differentially expressed were identified. Of the corresponding transcripts, 609 showed a significant sinusoidal expression pattern; about 40% of these exibithed a 24-hour periodicity while the other 60% was characterized by a shorter (about 12-hour) rhythm. We assigned the differentially expressed genes to functional categories and noticed that those concerning translation, proteolysis, energy and metabolic process, redox regulation, visual transduction and stress response, which are most likely related to daily environmental changes, were significantly enriched. Two transcripts of peroxiredoxin, thought to represent the ancestral timekeeping system that evolved about 2.5 billion years ago, were also identified as were two isoforms of the EsRh1 opsin and two novel arrestin1 sequences involved in the visual transduction cascade. CONCLUSIONS: Our work represents the first characterization of the krill diurnal transcriptome under natural conditions and provides a first insight into the genetic regulation of physiological changes, which occur around the clock during an Antarctic summer day Gene expression profiling was carried out in krill fished at different times throughout the 24 hours cycle (local times: 01:00, 06:00, 10:00, 15:00, and 18:00) with the M-bM-^@M-^XM-bM-^@M-^XKrill 1.1M-bM-^@M-^YM-bM-^@M-^Y custom platform (Agilent). We analyzed four different biological replicates for each time point for a total of 20 microarray experiments.

Dataset Information

Functional characterization of a Drosophila melanogaster mitochondrial deoxynucleotide transporter dRIM2.

Publications

Functional characterization of drim2, the Drosophila melanogaster homolog of the yeast mitochondrial deoxynucleotide transporter.

Similar Datasets

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