Biochemical characterization of a new mitochondrial transporter of dephosphocoenzyme A in Drosophila melanogaster.
CoA is an essential cofactor that holds a central role in cell metabolism. Although its biosynthetic pathway is conserved across the three domains of life, the subcellular localization of the eukaryotic biosynthetic enzymes and the mechanism behind the cytosolic and mitochondrial CoA pools compartmentalization are still under debate. In humans, the transport of CoA across the inner mitochondrial membrane has been ascribed to two related genes, SLC25A16 and SLC25A42 whereas in D. melanogaster genome only one gene is present, CG4241, phylogenetically closer to SLC25A42. CG4241 encodes two alternatively spliced isoforms, dPCoAC-A and dPCoAC-B. Both isoforms were expressed in Escherichia coli, but only dPCoAC-A was successfully reconstituted into liposomes, where transported dPCoA and, to a lesser extent, ADP and dADP but not CoA, which was a powerful competitive inhibitor. The expression of both isoforms in a Saccharomyces cerevisiae strain lacking the endogenous putative mitochondrial CoA carrier restored the growth on respiratory carbon sources and the mitochondrial levels of CoA. The results reported here and the proposed subcellular localization of some of the enzymes of the fruit fly CoA biosynthetic pathway, suggest that dPCoA may be synthesized and phosphorylated to CoA in the matrix, but it can also be transported by dPCoAC to the cytosol, where it may be phosphorylated to CoA by the monofunctional dPCoA kinase. Thus, dPCoAC may connect the cytosolic and mitochondrial reactions of the CoA biosynthetic pathway without allowing the two CoA pools to get in contact.
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: angelo.vozza@uniba.it.
Angelo
A
Vozza
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: francesco.deleonardis@uniba.it.
Francesco
F
De Leonardis
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy; CNR Institute of Biomembranes and Bioenergetics, via Amendola 165/A, 70126 Bari, Italy. Electronic address: e.paradies@ibbe.cnr.it.
Eleonora
E
Paradies
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: anna.degrassi@uniba.it.
Anna
A
De Grassi
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: ciroleopierri@gmail.com.
Ciro Leonardo
CL
Pierri
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: gvnparisi@gmail.com.
Giovanni
G
Parisi
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: carlo.marobbio@gmail.com.
Carlo Marya Thomas
CMT
Marobbio
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy; CNR Institute of Biomembranes and Bioenergetics, via Amendola 165/A, 70126 Bari, Italy. Electronic address: fmlasorsa@gmail.com.
Francesco Massimo
FM
Lasorsa
Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, 87036 Arcavacata di Rende, Cosenza, Italy. Electronic address: m.luigia@hotmail.it.
Luigina
L
Muto
Department of Biological and Environmental Sciences and Technologies, University of Salento, 73100 Lecce, Italy. Electronic address: loredana.capobianco@unisalento.it.
Loredana
L
Capobianco
Department of Pharmacy, Health and Nutritional Sciences, University of Calabria, 87036 Arcavacata di Rende, Cosenza, Italy. Electronic address: vincenza.dolce@unical.it.
Vincenza
V
Dolce
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: susanna.raho@gmail.com.
Susanna
S
Raho
Department of Biosciences, Biotechnologies and Biopharmaceutics, Laboratory of Biochemistry and Molecular Biology, University of Bari, via Orabona 4, 70125 Bari, Italy. Electronic address: giuseppe.fiermonte@uniba.it.
Giuseppe
G
Fiermonte
D000595:Amino Acid Sequence; D000818:Animals; D001692:Biological Transport / Q000502:physiology; D002352:Carrier Proteins / Q000378:metabolism; D003065:Coenzyme A / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004331:Drosophila melanogaster / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008928:Mitochondria / Q000378:metabolism*; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism*; D014176:Protein Biosynthesis / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D016415:Sequence Alignment
D002352:Carrier Proteins; D033681:Mitochondrial Membrane Transport Proteins; D029701:Saccharomyces cerevisiae Proteins; C015787:dephosphocoenzyme A; D003065:Coenzyme A
Bioenergetics; Coenzyme A; Dephosphocoenzyme A; Mitochondrial carrier; Neurodegenerative disease; Transport