Molecular cloning, functional expression and chromosomal localization of cdnas encoding a human na+/nucleoside cotransporter (hcnt1) . Ritzel, Mabel W. L., Sylvia Y. M. Yao, Mei-Yi Huang, John F. Elliott, Carol E. Cass, and James D. Young. Membrane Transport Group, Departments of Physiology, Biochemistry, Oncology and Medical Microbiology and Immunology, Medical Sciences Building, University of Alberta, Edmonton, Alberta, Canada T6G 2H7
APStracts 3:0315C, 1996.
We report identification of a new human nucleoside transporter protein by molecular cloning and functional expression of its cDNA. Previously, we used expression selection in Xenopus oocytes to isolate a cDNA from rat jejunal epithelium encoding the pyrimidine -selective Na+-dependent nucleoside transporter rCNT1 (Huang, Q.Q. et al (1994) J. Biol. Chem. 269, 17757-17760). cDNAs for a human homolog of rCNT1, designated hCNT1, have now been isolated from human kidney by hybridization cloning and reverse transcriptase polymerase chain reaction amplification strategies. hCNT1 was 83% identical to rCNT1 in amino acid sequence and exhibited the transport characteristics of a Na+-dependent nucleoside transporter with selectivity for pyrimidine nucleosides and adenosine when expressed in Xenopus oocytes. Deoxyadenosine, which undergoes net renal secretion, and guanosine were poor permeants. hCNT1 did, however, transport 3' -azido-3'-deoxythymidine. This is the first demonstration that members of the CNT family exist in human cells and provides evidence of their involvement in the renal transport of physiological nucleosides and nucleoside drugs. The hCNT1 gene was mapped to chromosome 15q25-26.

Received 14 May 1996; accepted in final form 23 September 1996.
APS Manuscript Number C263-6.
Article publication pending Am. J. Physiol. (Cell Physiology).
ISSN 1080-4757 Copyright 1996 The American Physiological Society.
Published in APStracts on 5 November 1996