title: The evolution of Jen3 proteins and their role in dicarboxylic acid transport in Yarrowia.
abstract: Jen proteins in yeast are involved in the uptake of mono/dicarboxylic acids. The Jen1 subfamily transports lactate and pyruvate, while the Jen2 subfamily transports fumarate, malate, and succinate. Yarrowia lipolytica has six JEN genes: YALI0B19470g, YALI0C15488g, YALI0C21406g, YALI0D20108g, YALI0D24607g, and YALI0E32901g. Through phylogenetic analyses, we found that these genes represent a new subfamily, Jen3 and that these three Jen subfamilies derivate from three putative ancestral genes. Reverse transcription-PCR. revealed that only four YLJEN genes are expressed and they are upregulated in the presence of lactate, pyruvate, fumarate, malate, and/or succinate, suggesting that they are able to transport these substrates. Analysis of deletion mutant strains revealed that Jen3 subfamily proteins transport fumarate, malate, and succinate. We found evidence that YALI0C15488 encodes the main transporter because its deletion was sufficient to strongly reduce or suppress growth in media containing fumarate, malate, or succinate. It appears that the other YLJEN genes play a minor role, with the exception of YALI0E32901g, which is important for malate uptake. However, the overexpression of each YLJEN gene in the sextuple-deletion mutant strain ΔYLjen1-6 revealed that all six genes are functional and have evolved to transport different substrates with varying degrees of efficacy. In addition, we found that YALI0E32901p transported succinate more efficiently in the presence of lactate or fumarate.
authors.affiliation: UMR1319 Micalis, INRA, Jouy-en-Josas, F-78352, France.
authors.forename: Rémi
authors.initials: R
authors.lastname: Dulermo
authors.forename: Heber
authors.initials: H
authors.lastname: Gamboa-Meléndez
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Michely
authors.forename: France
authors.initials: F
authors.lastname: Thevenieau
authors.forename: Cécile
authors.initials: C
authors.lastname: Neuvéglise
authors.forename: Jean-Marc
authors.initials: JM
authors.lastname: Nicaud
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D001692:Biological Transport; D029382:Dicarboxylic Acid Transporters / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019143:Evolution, Molecular; D005650:Fumarates / Q000378:metabolism; D005656:Fungal Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005800:Genes, Fungal; D008293:Malates / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D010802:Phylogeny; D017386:Sequence Homology, Amino Acid; D019802:Succinic Acid / Q000378:metabolism; D025062:Yarrowia / Q000235:genetics* / Q000378:metabolism
chemical_list: D029382:Dicarboxylic Acid Transporters; D005650:Fumarates; D005656:Fungal Proteins; D008293:Malates; C030298:malic acid; D019802:Succinic Acid
keywords: Fumarate; lactate; malate; succinate; transport; yeast