title: Genetic engineering and molecular characterization of yeast strain expressing hybrid human-yeast squalene synthase as a tool for anti-cholesterol drug assessment.
abstract: OBJECTIVE
  The main objective of the study is molecular and biological characterization of the human-yeast hybrid squalene synthase (SQS), as a promising target for treatment of hypercholesterolaemia.
  
  
  RESULTS
  The human-yeast hybrid SQS, with 67% amino acids, including the catalytic site derived from human enzyme, was expressed in Saccharomyces cerevisiae strain deleted of its own SQS gene. The constructed strain has a decreased level of sterols compared to the control strain. The mevalonate pathway and sterol biosynthesis genes are induced and the level of triacylglycerols is increased. Treatment of the strain with rosuvastatin or zaragozic acid, two mevalonate pathway inhibitors, decreased the amounts of squalene, lanosterol and ergosterol, and up-regulated expression of several genes encoding enzymes responsible for biosynthesis of ergosterol precursors. Conversely, expression of the majority genes implicated in the biosynthesis of other mevalonate pathway end products, ubiquinone and dolichol, was down-regulated.
  
  
  CONCLUSIONS
  The S. cerevisiae strain constructed in this study enables to investigate the physiological and molecular effects of inhibitors on cell functioning.
  
  
  CONCLUSIONS
  The yeast strain expressing hybrid SQS with the catalytic core of human enzyme is a convenient tool for efficient screening for novel inhibitors of cholesterol-lowering properties.
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: I
authors.initials: I
authors.lastname: Warchol
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Gora
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Wysocka-Kapcinska
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: J
authors.initials: J
authors.lastname: Komaszylo
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: E
authors.initials: E
authors.lastname: Swiezewska
authors.affiliation: Institute of Organic Chemistry, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Sojka
authors.affiliation: Institute of Organic Chemistry, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: W
authors.initials: W
authors.lastname: Danikiewicz
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Plochocka
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Maciejak
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: D
authors.initials: D
authors.lastname: Tulacz
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: A
authors.initials: A
authors.lastname: Leszczynska
authors.affiliation: Department of Biological Science, Birla Institute of Technology & Science (BITS), Hyderabad, India.
authors.forename: S
authors.initials: S
authors.lastname: Kapur
authors.affiliation: Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warsaw, Poland.
authors.forename: B
authors.initials: B
authors.lastname: Burzynska
mesh_terms: D000924:Anticholesteremic Agents / Q000378:metabolism*; D002784:Cholesterol / Q000378:metabolism*; D004875:Ergosterol / Q000378:metabolism; D013186:Farnesyl-Diphosphate Farnesyltransferase / Q000235:genetics / Q000378:metabolism*; D005818:Genetic Engineering; D006801:Humans; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D013185:Squalene / Q000378:metabolism; D014280:Triglycerides / Q000378:metabolism; D015854:Up-Regulation
chemical_list: D000924:Anticholesteremic Agents; D011993:Recombinant Fusion Proteins; D014280:Triglycerides; D013185:Squalene; D002784:Cholesterol; D013186:Farnesyl-Diphosphate Farnesyltransferase; D004875:Ergosterol
keywords: heterologous proteins; mevalonate pathway; squalene synthase; sterol biosynthesis inhibitors; yeast expression system