Two mRNA transcripts (rBAT-1 and rBAT-2) are involved in system b0,(+)-related amino acid transport.
Previously, we isolated a cDNA clone (rBAT-1) of 2.2 kilobase pairs (kb) from a rabbit kidney cortex cDNA library, encoding a protein involved in sodium-independent transport of L-dibasic amino acids, L-cystine, and some neutral amino acids via a system related to b0,(+)-like activity (Bertran, J., Werner, A., Moore, M. L., Stange, G., Markovich, D., Biber, J., Testar, X., Zorzano, A., Palacin, M., and Murer, H. (1992) Proc. Natl. Acad. Sci. U. S. A. 89, 5601-5605). In Northern blot hybridization using an rBAT-1 cDNA probe, 2.2- and 3.9-kb mRNA species were observed. Here we describe the isolation of a 3.9-kb cDNA clone (rBAT-2) by expression cloning using Xenopus laevis oocytes corresponding to the 3.9-kb mRNA species. On the basis of sequence analysis, in vitro translation (major protein of approximately 78 kDa), and functional analysis (expression of transport function), we conclude that rBAT-1- and rBAT-2-related proteins are identical: 677 amino acids in length, with most likely only one transmembrane-spanning domain. There are seven differences in the nucleotide composition within a common overlap of 2189 nucleotides, resulting in 2 amino acid replacements. In comparison with rBAT-1, rBAT-2 has 26 additional nucleotides at the 5'-end, an identical location of the first polyadenylation signal, and approximately 1.7 kb of 3'-untranslated sequence (rich in AT(U) motifs) prior to a poly(A) tail of 63 adenines. We conclude that rBAT-1 and rBAT-2 encode the same protein and that the major difference seems to be related to the use of different polyadenylation signals.
Markovich
D
D
Department of Physiology, University of Zürich, Switzerland.
Stange
G
G
Bertran
J
J
Palacin
M
M
Werner
A
A
Biber
J
J
Murer
H
H
D000595:Amino Acid Sequence; D026923:Amino Acid Transport Systems, Basic; D000599:Amino Acids, Diamino / Q000378:metabolism*; D000818:Animals; D001120:Arginine / Q000378:metabolism; D001483:Base Sequence; D001692:Biological Transport; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D003553:Cystine / Q000378:metabolism; D005260:Female; D006714:Homoserine / Q000494:pharmacology; D007672:Kidney Cortex / Q000378:metabolism*; D007930:Leucine / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D011817:Rabbits; D012689:Sequence Homology, Nucleic Acid; D014158:Transcription, Genetic; D014982:Xenopus laevis
D026923:Amino Acid Transport Systems, Basic; D000599:Amino Acids, Diamino; D002352:Carrier Proteins; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; C082087:rBAT protein, Oryctolagus cuniculus; D003553:Cystine; D006714:Homoserine; D001120:Arginine; D007930:Leucine