High affinity copper transport protein in the lizard Podarcis sicula: Molecular cloning, functional characterization and expression in somatic tissues, follicular oocytes and eggs

Marilisa Riggio, Jaekwon Lee, Rosaria Scudiero, Elio Parisi, Dennis J. Thiele, Silvana Filosa

Research output: Contribution to journalArticle

22 Scopus citations

Abstract

Copper (Cu) is an essential element required in many biological processes including cellular growth and development. The molecular mechanisms involved in copper homeostasis include proteins that play a role in Cu uptake. Genes encoding high affinity copper transporters (Ctr) have been identified in yeast, plant and mammalian cells. Analysis of copper and zinc content in growing ovarian follicles and ovulated eggs of the reptilian Podarcis sicula demonstrated that the levels of both metals rise during oocyte growth, reaching the maximum in ovulated eggs. By exploiting the remarkable evolutionary conservation of the primary structure of Ctr proteins, cDNA encoding a Ctr was isolated from the liver of the lizard P. sicula by reverse transcriptase PCR and RACE strategy by using primers designed based on consensus motifs present in mammalian Ctr. The predicted protein sequence contains three transmembrane domains and a putative hydrophilic extracellular amino-terminal domain. Besides complementing the respiratory deficiency of yeast cells defective in high affinity Cu transport, expression of lizard Ctr11 in Hek293 cells stimulates Cu uptake. Gene expression assessed by Northern blot hybridization of RNA from different tissues of P. sicula shows the highest levels of transcript in both intestine and liver. The profile of Ctr1 mRNA in growing ovarian follicles and eggs demonstrates that the transcript accumulates during the oocyte growth and reaches the highest levels in ovulated eggs. These results suggest that lizard Ctr1 protein may function in Cu acquisition in growing oocytes and eggs.

Original languageEnglish (US)
Pages (from-to)127-135
Number of pages9
JournalBiochimica et Biophysica Acta - Gene Structure and Expression
Volume1576
Issue number1-2
DOIs
StatePublished - Jun 7 2002

Keywords

  • Copper
  • Copper transporter
  • Lizard
  • Oogenesis
  • Zinc

ASJC Scopus subject areas

  • Structural Biology
  • Biophysics
  • Biochemistry
  • Genetics

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