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Information × Registration Number 0213U003135, 0110U006325 , R & D reports Title Studying the selective autophagic degradation of organelles and cytosolic proteins in yeasts Pichia pastoris and Saccharomyces cerevisiae popup.stage_title Head Sibirny Andriy Andriyovych, Registration Date 24-01-2013 Organization Institute of cell biology, National Academy of Sciences of Ukraine popup.description2 On this stage of work it was shown that FBP P. pastoris and H. polymorpha is mostly degraded after 5 hours of incubation of cells, pregrown in gluconeogenic substrates, in glucose-containing medium. After shifting the cells from methanol or ethanol-containing media on glucose medium, FBPase activity of P. pastoris and H. polymorpha wild-type strains decreased 4,5-5 and 2-2,5 times, respectively. The recombinant strains of P. pastoris with the deletion of FBP gene were constructed and analyzed. DeltaFBP strains did not grow in the medium with non-fermentable carbon sources, such as methanol and ethanol, and revealed impaired growth in glycerol-containing medium. To study the mechanisms of catabolite degradation, plasmids and strains with the expression of FBP fused with GFP under the control of FBP prompter were constructed and analysed. The FBPase activity of obtained transformants was in 4 times less that the activity of wild-type strain. It was shown that the deletion of HXK1, GLK1 and HXK2 genes in S. cerevisiae has no effect on pexophagy. The deletion of both hexokinases (HXK1and HXK2) or hexokinase 2 with the absence of glucokinase 1 (GLK1and HXK1) results in slower growth of these strains on the medium with oleate and decreased the level of tiolase in the cells of such mutants. The mutation in regulatory domain and the deletion of HXK2 gene had no effect on the biogenesis of peroxisomes . The mutation in the hexokinase 2 gene had no effect on aminopeptidase maturation and did not damage the Cvt-pathway. The damage of HXK2wrf and HXK2wca domaines of hexokinase 2 had no effect on nonspecific macroautophagy.5481 Product Description popup.authors Назарко Володмир Юрійович Полупанов Андрій Сергійович Сибірний Андрій Андрійович popup.nrat_date 2020-04-02 Close