Subcellular localization of oxidosqualene cyclases from Arabidopsis thaliana, Trypanosoma cruzi, and Pneumocystis carinii expressed in yeast

  title={Subcellular localization of oxidosqualene cyclases from Arabidopsis thaliana, Trypanosoma cruzi, and Pneumocystis carinii expressed in yeast},
  author={Paola Milla and Franca Viola and Simonetta Oliaro Bosso and Flavio Rocco and Luigi Cattel and Bridget M. Joubert and Ren{\'e}e J Leclair and Seiichi P T Matsuda and Gianni Balliano},
Cycloartenol synthase from Arabidopsis thaliana and lanosterol synthase from Trypanosoma cruzi and Pneumocystis carinii were expressed in yeast, and their subcellular distribution in the expressing cells was compared. Determination of enzymatic (oxidosqualene cyclase, OSC) activity and SDS-PAGE analysis of subcellular fractions proved that enzymes from T. cruzi and A. thaliana have high affinity for lipid particles, a subcellular compartment rich in triacylglycerols, and steryl esters… 
Inhibitory Effect of Umbelliferone Aminoalkyl Derivatives on Oxidosqualene Cyclases from S. cerevisiae, T. cruzi, P. carinii, H. sapiens, and A. thaliana: a Structure–Activity Study
Eighteen coumarin derivatives were tested as inhibitors of oxidosqualene cyclases from Saccharomyces cerevisiae, Trypanosoma cruzi, Pneumocystis carinii, Homo sapiens, and Arabidopsis thaliana and suggest the present series as a promising compound family for the development of novel antiparasitic agents.
Functional Characterization and Localization of Pneumocystis carinii Lanosterol Synthase
Using fluorescence microscopy, it is shown for the first time the presence of apparent lipid particles in P. carinii and the localization of PcErg7p to cholesterol particles in yeast, providing strong evidence that the enzyme serves a similar function in the P.Carinii sterol pathway.
Analogs of squalene and oxidosqualene inhibit oxidosqualene cyclase of Trypanosoma cruzi expressed in Saccharomyces cerevisiae
The expression in yeast cells could be a safe and easy model for studying the activity and the selectivity of the potential inhibitors of T. cruzi OSC, and the expression of proteins of pathogenic organisms in yeast seems very promising for preliminary screening of compounds that have potential therapeutic activity.
Umbelliferone aminoalkyl derivatives as inhibitors of oxidosqualene cyclases from Saccharomyces cerevisiae, Tripanosoma cruzi, and Pneumocystis carinii
Results identify a particularly promising new family of OSC inhibitors, for the development of novel antiparasitic agents, including derivatives bearing a methylallylaminoalkyl flexible chain.
Difference in the late ergosterol biosynthesis between yeast spheroplasts and intact cells.
A comparative study on post-squalene sterol synthesis in intact yeast cells and spheroplasts was carried out with strains from three genera as well as with engineered S. cerevisiae cells altered in regard to the late ergosterol synthesis pathway to search for an answer.
Pneumocystis carinii Sterol 14α‐Demethylase Activity in Saccharomyces cerevisiae erg11 Knockout Mutant: Sterol Biochemistry
Detailed sterol analysis of wild‐type Saccharomyces cerevisiae and in an erg11 knockout mutant expressing either P. carinii or S. Cerevisiae 14DM indicated that the mutant transformed with erg11 showed more 14DM activity than wild‐ type yeast and suggested that the yeast in the yeast cell utilizes 4α‐methyl sterols better than the P.carinii enzyme.
Plant Oxidosqualene Metabolism: Cycloartenol Synthase–Dependent Sterol Biosynthesis in Nicotiana benthamiana
A CAS1-specific functional sterol pathway in engineered yeast, and a strict dependence on CAS1 of tobacco sterol biosynthesis are shown.
Sterol biosynthesis and sterol uptake in the fungal pathogen Pneumocystis carinii.
Members of the fungal genus Pneumocystis colonize healthy mammalian hosts without causing apparent disease, but colonization in immunocompromised hosts may result in a potentially fatal pneumonia
Identification of marneral synthase, which is critical for growth and development in Arabidopsis.
  • Y. Go, S. Lee, M. Suh
  • Environmental Science
    The Plant journal : for cell and molecular biology
  • 2012
Evidence is provided that the unusual triterpenoid pathway via marneral synthase is important for the growth and development of Arabidopsis.
Enfumafungin synthase represents a novel lineage of fungal triterpene cyclases
The preliminary identification of the enfumafungin biosynthetic gene cluster (BGC) is reported based on genome sequencing, phylogenetic reconstruction, gene disruption, and cDNA sequencing studies, suggesting that efuA represents the first member of a widespread lineage of fungal SHCs.


Yeast Oxidosqualene Cyclase (Erg7p) Is a Major Component of Lipid Particles*
Evidence is provided that lipid particles are not only a depot for neutral lipids, but also participate in coordinate sterol metabolism and trafficking and serve as a storage site for compounds that may negatively affect membrane integrity.
Isolation of an Arabidopsis thaliana gene encoding cycloartenol synthase by functional expression in a yeast mutant lacking lanosterol synthase by the use of a chromatographic screen.
The cloning and characterization of CAS1, an Arabidopsis thaliana gene encoding cycloartenol synthase, is reported, which should be generally applicable to genes responsible for secondary metabolite biosynthesis.
Dual localization of squalene epoxidase, Erg1p, in yeast reflects a relationship between the endoplasmic reticulum and lipid particles.
Analysis of subcellular fractions revealed that squalene epoxidase was present in the microsomal fraction and also cofractionated with lipid particles, suggesting that close contact between lipid particles and endoplasmic reticulum may be necessary for a concerted action of these two compartments in sterol biosynthesis.
Structure and function of a squalene cyclase.
The crystal structure of squalene-hopene cyclase from Alicyclobacillus acidocaldarius was determined and reveals a membrane protein with membrane-binding characteristics similar to those of prostaglandin-H2 synthase, the only other reported protein of this type.
Characterization of lipid particles of the yeast, Saccharomyces cerevisiae
In lipid particles of the yeast mutant strain S. cerevisiae erg6, this protein is missing thereby identifying the protein and confirming the previous finding (Zinser et al., 1993) that sterol Δ24‐methylation is associated with lipid particles.
Contribution of Are1p and Are2p to steryl ester synthesis in the yeast Saccharomyces cerevisiae.
In a growth competition experiment are1are2 cells grow more slowly than wild-type after several rounds of cultivation, suggesting that Are1p and Are2p or steryl esters, the product formed by these two enzymes, are more important in the natural environment than under laboratory conditions.
Molecular cloning of a Schizosaccharomyces pombe cDNA encoding lanosterol synthase and investigation of conserved tryptophan residues.
All of the resultant mutant enzymes retained the ability to complement the Sc.
The structure of the membrane protein squalene-hopene cyclase at 2.0 A resolution.
The structure of the enzyme from Alicyclobacillus acidocaldarius has been determined and the membrane-binding interactions of which are described and compared with those of two prostaglandin-H2 synthase isoenzymes, the only other structurally characterized proteins of this type.