Identification of the Drosophila melanogaster mitochondrial citrate carrier: bacterial expression, reconstitution, functional characterization and developmental distribution.
The mitochondrial carriers are a family of transport proteins that shuttle metabolites, nucleotides and cofactors across the inner mitochondrial membrane. The genome of Drosophila melanogaster encodes at least 46 members of this family. Only four of them have been characterized: the two isoforms of the ADP/ATP translocase, the brain uncoupling protein and the carnitine/acylcarnitine carriers. The transport functions of the remainders cannot be assessed with certainty. One of them, the product of the gene CG6782, shows a fairly close sequence homology to the known sequence of the rat mitochondrial citrate carrier. In this article the fruit fly protein coding by the CG6782 gene has been functionally characterized by over-expression in Escherichia coli and reconstitution into liposomes. It shows to have similar transport properties of the eukaryotic mitochondrial citrate carriers previously biochemically characterized. This indicates that in addition to the protein sequence conservation, insect and mammalian citrate carriers are also significantly related at the functional level suggesting that Drosophila may be used as model organism for the study of mitochondrial solute transporter. The DmCIC expression pattern throughout development was also investigated; the transcripts were detected at equal levels in all stages analysed.
Department of Biological and Environmental Sciences, University of Salento, Lecce, Italy.
Chiara
C
Carrisi
Marianna
M
Madeo
Patrizia
P
Morciano
Vincenza
V
Dolce
Giovanni
G
Cenci
Anna Rita
AR
Cappello
Giancarlo
G
Mazzeo
Domenico
D
Iacopetta
Loredana
L
Capobianco
D000818:Animals; D002352:Carrier Proteins / Q000737:chemistry* / Q000502:physiology*; D002951:Citrates / Q000737:chemistry*; D004331:Drosophila melanogaster / Q000196:embryology / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D005786:Gene Expression Regulation; D008081:Liposomes / Q000737:chemistry; D008928:Mitochondria / Q000378:metabolism*; D008954:Models, Biological; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D012260:Ribonucleases / Q000737:chemistry; D013997:Time Factors; D014018:Tissue Distribution
D002352:Carrier Proteins; D002951:Citrates; D008081:Liposomes; D011994:Recombinant Proteins; C020723:citrate-binding transport protein; D012260:Ribonucleases