Submitted on May 15, 2006
Accepted on October 27, 2006
Genomic analysis of nucleoside transporters in Diptera and functional characterization of DmENT2, a Drosophila equilibrative nucleoside transporter
Jerry Machado1, Parween Abdulla1, WJ Brad Hanna2, Arthur J Hilliker1, and Imogen R Coe1*
1 Biology, York University, Toronto, Canada
2 Biomedical Sciences, University of Guelph, Guelph, Canada
* To whom correspondence should be addressed. E-mail: coe{at}yorku.ca.
The recent completion of genome sequencing projects in a number of eukaryotes allows comparative analysis of orthologs, which can aid in identifying evolutionary constraints on protein structure and function. Nucleoside transporters (NTs) are present in a diverse array of organisms and previous studies have suggested that there is low protein sequence similarity but conserved structure in invertebrate and vertebrate NT orthologs. In addition, most taxa possess multiple NT isoforms but their respective roles in the physiology of the organism are not clear. To investigate the evolution of the structure and function of NTs, we have extended our previous studies by identifying NT orthologs in the Dipteran, Anopheles gambiae and comparing these proteins to human and Drosophila melanogaster NTs. In addition, we have functionally characterized DmENT2, one of three putative Drosophila melanogaster ENTs that we have previously described. DmENT2 has broad substrate specificity, is insensitive to standard nucleoside transport inhibitors and is expressed in the digestive tract of late stage embryos based on in situ hybridization. DmENT1 and DmENT2 are expressed in most stages during development with the exception of early embryogenesis suggesting specific physiological roles for each isoform. These data represent the first complete genomic analysis of Dipteran NTs and the first report of the functional characterization of any Dipteran NT.
