Molecular cloning and expression of human xCT, the light chain of amino acid transport system xc-.
Transport of system xc- is an exchange agency with high specificity for anionic form of cystine and glutamate. The protein mediating this transport is a disulfide-linked heterodimer of a light chain named xCT and a heavy chain previously known as 4F2hc. We have isolated two cDNAs encoding xCT from the human cDNA library. One clone coded for a protein of 501 amino acids with 12 putative transmembrane domains. When functionally expressed in Xenopus oocytes together with the human 4F2hc, human xCT induced the transport activity whose characteristics are similar to those of system xc-. Another clone seemed to contain a partial human xCT and a long 3' untranslated region. The human xCT gene was localized at chromosome 4q28-31. Analysis of the 5'-flanking region of the human xCT gene revealed several sites for potentially binding of transcriptional factors, including NF-E2 and AP-1. Transport of cystine via system xc- has been known as a regulatory factor for the intracellular glutathione level, and its transport activity is induced in response to the oxygen tension in culture. Northern blot analysis demonstrated that the expression of both xCT and 4F2hc was significantly enhanced by oxygen. The results suggest that oxygen regulates the activity of system xc- by modulating the expression of both xCT and 4F2hc mRNAs.
Sato
H
H
Department of Biochemistry, Institute of Basic Medical Sciences, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
Tamba
M
M
Kuriyama-Matsumura
K
K
Okuno
S
S
Bannai
S
S
D000595:Amino Acid Sequence; D027182:Amino Acid Transport System y+; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D002461:Cell Line, Transformed; D002874:Chromosome Mapping; D002894:Chromosomes, Human, Pair 4 / Q000235:genetics; D003001:Cloning, Molecular; D003553:Cystine / Q000378:metabolism; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics / Q000302:isolation & purification; D005260:Female; D015870:Gene Expression; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014981:Xenopus
D027182:Amino Acid Transport System y+; D002352:Carrier Proteins; D017931:DNA Primers; D018076:DNA, Complementary; D011994:Recombinant Proteins; C118895:SLC7A11 protein, human; C492255:Slc7a11 protein, mouse; D018698:Glutamic Acid; D003553:Cystine