Molecular characterization and functional analysis of Cashmere goat mammalian target of rapamycin

DNA Cell Biol. 2012 May;31(5):839-44. doi: 10.1089/dna.2011.1393. Epub 2011 Dec 16.

Abstract

The mammalian target of rapamycin (mTOR) is an evolutionarily conserved protein kinase that belongs to the phosphatidylinositol kinase-related kinase family. We describe our molecular characterization of mTOR and its function (GenBank accession HM114224) in Cashmere goat (Capra hircus). The goat mTOR complementary DNA is 8617 bp, comprising an open reading frame of 7650 bp--corresponding to a polypeptide of 2549 amino acids--and a 909 bp 3' untranslated region with a polyA tract and a polyadenylation signal at nucleotides 8575-8580. In a bioinformatics analysis, goat mTOR has typical sites of activity and domains. mTOR mRNA was measured in brain, heart, testis, liver, spleen, kidney, and lung by real-time polymerase chain reaction, and the expression of mTOR in fetal fibroblasts was detected by western blot. The viability of fetal fibroblasts was inhibited on treatment with CCI-779, a specific inhibitor of mTOR. Our data supplied evidence that the transcription of mTOR was detected in the seven tissues in Cashmere goat, and mTOR protein was translated in fetal fibroblasts. The proliferation of fetal fibroblasts decreases on inhibition of mTOR.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Blotting, Western
  • Cell Proliferation / drug effects
  • Cells, Cultured
  • Cloning, Molecular
  • Fetus / cytology
  • Fetus / drug effects
  • Fetus / metabolism*
  • Fibroblasts / cytology
  • Fibroblasts / drug effects
  • Fibroblasts / metabolism*
  • Goats
  • Models, Molecular
  • Molecular Sequence Data
  • Protein Conformation
  • Protein Kinase Inhibitors / pharmacology
  • RNA, Messenger / genetics
  • Real-Time Polymerase Chain Reaction
  • Sequence Homology, Amino Acid
  • Sirolimus / analogs & derivatives
  • Sirolimus / pharmacology
  • TOR Serine-Threonine Kinases / antagonists & inhibitors
  • TOR Serine-Threonine Kinases / genetics*
  • TOR Serine-Threonine Kinases / metabolism*
  • Tissue Distribution

Substances

  • Protein Kinase Inhibitors
  • RNA, Messenger
  • temsirolimus
  • TOR Serine-Threonine Kinases
  • Sirolimus