Functional redundancy of two nucleoside transporters of the ENT family (CeENT1, CeENT2) required for development of Caenorhabditis elegans.
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.
Appleford
Peter J
PJ
School of Biochemistry & Microbiology, University of Leeds, Leeds LS2 9JT, UK.
Griffiths
Mark
M
Yao
Sylvia Y M
SY
Ng
Amy M L
AM
Chomey
Eugene G
EG
Isaac
R Elwyn
RE
Coates
David
D
Hope
Ian A
IA
Cass
Carol E
CE
Young
James D
JD
Baldwin
Stephen A
SA
D000595:Amino Acid Sequence; D000818:Animals; D017173:Caenorhabditis elegans / Q000196:embryology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D029742:Caenorhabditis elegans Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D033721:Equilibrative Nucleoside Transporter 1; D033722:Equilibrative-Nucleoside Transporter 2; D007700:Kinetics; D008969:Molecular Sequence Data; D033703:Nucleoside Transport Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000187:drug effects / Q000378:metabolism; D010802:Phylogeny; D034622:RNA Interference; D016415:Sequence Alignment; D014529:Uridine / Q000494:pharmacology; D014982:Xenopus laevis / Q000235:genetics / Q000378:metabolism
D029742:Caenorhabditis elegans Proteins; D033721:Equilibrative Nucleoside Transporter 1; D033722:Equilibrative-Nucleoside Transporter 2; D033703:Nucleoside Transport Proteins; C500173:equilibrative nucleoside transporter 1, C elegans; C500174:equilibrative nucleoside transporter 2, C elegans; D014529:Uridine