Isolation of a murine copper transporter gene, tissue specific expression and functional complementation of a yeast copper transport mutant

A polymerase chain reaction (PCR)-based strategy was used to isolate a mous e cDNA (mCtr1) encoding a Cu transport protein. The deduced mCtr1 protein s equence exhibits 92% identity to human Ctr1, and has structural features in common with known high affinity Cu transporters from yeast. The expression of mouse Ctr1 functionally complements baker's yeast cells defective in hi gh affinity Cu transport. Characterization of the mCtr1 genomic clone showe d that the mCtr1 coding sequence is encompassed within four exons and that the mCtr1 locus maps to chromosome band 4Cl-2. RNA blotting analysis demons trated that mCtr1 is ubiquitously expressed, with high levels in liver and kidney, and early in embryonic development. Steady state mammalian Ctr1 mRN A levels were not changed in response to cellular Cu availability, which is distinct from the highly Cu-regulated transcription of genes encoding yeas t high affinity Cu transporters. These studies provide fundamental informat ion for further investigations on the function and regulation of Ctr1 in Cu acquisition in mammals. (C) 2000 Elsevier Science B.V. All rights reserved .