Cloning and Characterization of Human NTT5 and v7-3: Two Orphan Transporters of the Na+/Cl−-Dependent Neurotransmitter Transporter Gene Family

doi:10.1006/geno.2000.6387

Genomics

Volume 70, Issue 2, 1 December 2000, Pages 241-252

https://doi.org/10.1006/geno.2000.6387 Get rights and content

Abstract

Orphan transporters form a growing subfamily of genes related by sequence similarity to the Na⁺/Cl⁻-dependent neurotransmitter superfamily. Using a combination of database similarity searching and cloning methods, we have identified and characterized two novel human orphan transporter genes, v7-3 and NTT5. Similar to other known orphan transporters, v7-3 and NTT5 contain 12 predicted transmembrane domains, intracellular N- and C-terminal domains, and large extracellular loops between transmembrane (TM) domains 3 and 4 and between TM domains 7 and 8. Residues within the extracellular loops are also predicted to contain sites for N-linked glycosylation. Human v7-3, the species orthologue of rat v7-3, contains an open reading frame (ORF) of 730 amino acids. Human NTT5 is a new member of the orphan transporter family and has an ORF of 736 amino acids. The amino acid sequences of human v7-3 and NTT5 are greater than 50% similar to other known orphan neurotransmitter transporters and also show sequence similarity to the human serotonin and dopamine transporters. Radiation hybrid mapping located the human v7-3 and NTT5 genes on chromosomes 12q21.3–q21.4 and 19q13.1–q13.4, respectively. Human mRNA distribution analysis by TaqMan reverse transcription-polymerase chain reaction showed that v7-3 mRNA is predominantly expressed in neuronal tissues, particularly amygdala, putamen, and corpus callosum, with low-level expression in peripheral tissues. In contrast, NTT5 mRNA was highly expressed in peripheral tissues, particularly in testis, pancreas, and prostate. Transient transfection with epitope-tagged transporter constructs demonstrated v7-3 to be expressed at the cell surface, whereas NTT5 was predominantly intracellular, suggestive of a vesicular location. Although the substrates transported by these transporters remain unknown, their specific but widespread distribution suggests that they may mediate distinct and important functions within the brain and the periphery.

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^☆: Sequence data from this article have been deposited with the EMBL/GenBank Data Libraries under Accession Nos. AF265577 (human v7-3) and AF265578 (human NTT5).

¹: To whom correspondence should be addressed. Telephone: +44 1279 622971. Fax: +44 1279 622230. E-mail: [email protected].

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Regular ArticleCloning and Characterization of Human NTT5 and v7-3: Two Orphan Transporters of the Na+/Cl−-Dependent Neurotransmitter Transporter Gene Family☆

Abstract

Curr. Opin. Neurobiol.

J. Biol. Chem.

J. Biol. Chem.

Mol. Brain Res.

Eur. J. Pharmacol.

J. Biol. Chem.

FEBS Lett.

Genet. Anal.

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J. Biol. Chem.

J. Biol. Chem.

FEBS Lett.

FEBS Lett.

FEBS Lett.

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Regular Article
Cloning and Characterization of Human NTT5 and v7-3: Two Orphan Transporters of the Na⁺/Cl⁻-Dependent Neurotransmitter Transporter Gene Family☆