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Abstract
The genome of Caenorhabditis elegans encodes multiple homologues of the two major families of mammalian equilibrative and concentrative nucleoside transporters. As part of a programme aimed at understanding the biological rationale underlying the multiplicity of eukaryote nucleoside transporters, we have now demonstrated that the nematode genes ZK809.4 (ent-1) and K09A9.3 (ent-2) encode equilibrative transporters, which we designate CeENT1 and CeENT2 respectively. These transporters resemble their human counterparts hENT1 and hENT2 in exhibiting similar broad permeant specificities for nucleosides, while differing in their permeant selectivities for nucleobases. They are insensitive to the classic inhibitors of mammalian nucleoside transport, nitrobenzylthioinosine, dilazep and draflazine, but are inhibited by the vasoactive drug dipyridamole. Use of green fluorescent protein reporter constructs indicated that the transporters are present in a limited number of locations in the adult, including intestine and pharynx. Their potential roles in these tissues were explored by using RNA interference to disrupt gene expression. Although disruption of ent-1 or ent-2 expression alone had no effect, simultaneous disruption of both genes yielded pronounced developmental defects involving the intestine and vulva.
bp, base pair(s); BLAST, basic local alignment search tool; Ce, Caenorhabditis elegans (e.g. in CeENT1); CNT, concentrative nucleoside transporter; dbEST, expressed sequence tag database; dsRNA, double-stranded RNA; ei, equilibrative nitrobenzylthioinosine-insensitive; ENT, equilibrative nucleoside transporter; es, equilibrative nitrobenzylthioinosine-sensitive; EST, expressed sequence tag; GFP, green fluorescent protein; NBMPR, nitrobenzylthioinosine (6-[(4-nitrobenzyl)thio]-9-β- -ribofuranosylpurine); PCR, polymerase chain reaction; Pwo, Pyrococcus woesei; RNAi, RNA interference; TM, transmembrane.
bp, base pair(s); BLAST, basic local alignment search tool; Ce, Caenorhabditis elegans (e.g. in CeENT1); CNT, concentrative nucleoside transporter; dbEST, expressed sequence tag database; dsRNA, double-stranded RNA; ei, equilibrative nitrobenzylthioinosine-insensitive; ENT, equilibrative nucleoside transporter; es, equilibrative nitrobenzylthioinosine-sensitive; EST, expressed sequence tag; GFP, green fluorescent protein; NBMPR, nitrobenzylthioinosine (6-[(4-nitrobenzyl)thio]-9-β- -ribofuranosylpurine); PCR, polymerase chain reaction; Pwo, Pyrococcus woesei; RNAi, RNA interference; TM, transmembrane.