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Projects / Programmes source: ARIS

Lanosterol 14alpha-demethylase in biosynthesis of cholesterol and signalling sterols

Research activity

Code Science Field Subfield
1.05.00  Natural sciences and mathematics  Biochemistry and molecular biology   

Code Science Field
P004  Natural sciences and mathematics  Biochemistry, Metabolism 
P320  Natural sciences and mathematics  Nucleic acids, protein synthesis 
P340  Natural sciences and mathematics  Lipids, steroids, membranes 
Keywords
lanosterol 14alpha-demethylase, cytochrome P450, gene expression and regulation, transcriptome analysis, DNA chip technology, functional genomics, cholesterol biosynthesis, MAS sterols
Evaluation (rules)
source: COBISS
Researchers (12)
no. Code Name and surname Research area Role Period No. of publicationsNo. of publications
1.  17572  PhD Marko Cotman  Veterinarian medicine  Researcher  2004 - 2007  119 
2.  18622  PhD Nataša Debeljak  Biochemistry and molecular biology  Researcher  2004 - 2007  249 
3.  18992  PhD Martina Fink  Biochemistry and molecular biology  Researcher  2004 - 2007  128 
4.  20347  PhD Klementina Fon Tacer  Metabolic and hormonal disorders  Junior researcher  2004 - 2005  126 
5.  12449  PhD Robert Frangež  Veterinarian medicine  Researcher  2004 - 2007  280 
6.  24348  PhD Neža Grgurevič  Veterinarian medicine  Junior researcher  2004 - 2007  58 
7.  24562  Helena Klavžar  Biochemistry and molecular biology  Technical associate  2004 - 2007 
8.  18355  PhD Drago Kuzman  Physics  Researcher  2004 - 2007  64 
9.  13330  PhD Gregor Majdič  Veterinarian medicine  Researcher  2004 - 2007  579 
10.  22459  PhD Tadeja Režen  Neurobiology  Junior researcher  2004 - 2007  242 
11.  06013  PhD Damjana Rozman  Biochemistry and molecular biology  Head  2004 - 2007  888 
12.  24289  PhD Matej Seliškar  Biochemistry and molecular biology  Junior researcher  2004 - 2007  25 
Organisations (3)
no. Code Research organisation City Registration number No. of publicationsNo. of publications
1.  0258  Lek Pharmaceutical Company d.d.  Ljubljana  1732811  8,210 
2.  0381  University of Ljubljana, Faculty of Medicine  Ljubljana  1627066  48,407 
3.  0406  University of Ljubljana, Veterinary Faculty  Ljubljana  1627139  10,800 
Abstract
Disturbed cholesterol homeostasis leads to hyperlipidemias that represent one of the major causes of death in developed societies. The preceeding project contributed importantly to understanding the tissue-specific mechanisms of cholesterol and meiosis activating sterol (MAS) synthesis, at the level of gene regulation (trans-activation of lanosterol 14alpha demethylase CYP51 in sterol-repressed conditions), protein expression (CYP51 enzymatic activity in acrosomal membranes of sperm) and intracellular transport (traffick from ER through the Golgi to the acrosome). The vision of this project is to translate the knowledge of the multi-level regulation of CYP51 to other genes of cholesterol homeostasis (the STEROLTALK group), by applying functional genomic tools in studies of the cross-talk of cholesterol homeostasis with endogenous and exogenous pathways. Due to a high evolutionarily conservation of cholesterol synthesis and metabolism in mammals, several animal models will be applied, from mouse (in vivo studies) to big mammals (bull and ram), in addition to human immortal cell lines. Regulation of cholesterogenic genes will be studied, with particular emphasis on tissue-sepcificity and signalling pathways, that interact with the cholesterol feedback loop. By applying expression profiling by DNA chips and real time PCR and promoter studies, the physiological and patophysiological conditions that allow a cholesterol-feedback independent expression will be identified. The amount of cholesterol and MAS will be measured in animal tissues and cell cultures where different signalling pathways will be activated. Methods for proteome studies will be applied to determine the composition of CYP51 protein compelxes in Golgi and on the acrosome. The CYP51 enzyme activity will be measured in different subcellular fractions and the intracellular transport of CYP51-GFP-fusion protein evaluated by confocal microscopy. Factors that prevent the homologous recombination of the cyp51-lox P constructs into the mouse genome will be determined and new recombination constructs prepared, contributing to the assession of the final goal - a cyp51 conditional knockout mouse by the Cre-loxP technology.
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