Search academic texts

Information × Registration Number 0221U107074, 0121U113776 , R & D reports Title Identification of genes involved in autophagic degradation of soluble proteins in yeast and human cells popup.stage_title Head Sibirny Andriy A., Доктор біологічних наук Registration Date 26-12-2021 Organization Institute of Cell Biology of the National Academy of Sciences of Ukraine popup.description2 A system for selection of recombinant strains of Komagatella phaffii with disrupted genes associated with autophagic degradation of cytosolic proteins has been developed. For that, the Kluyveromyces lactis gene LAC4, encoding β-galactosidase, was expressed under the control of methanol-induced promoter of the FLD1 gene in K. phaffii strain GS200. Based on the constructed strain GS200/LAC4, insertion mutants were obtained in which the process of β-galactosidase degradation was slowed down compared to the original strain. In one strain, the insertion cassette integration site was identified - the integration occurred in the region of ​​the ORF of the gene encoding β-1,6-N-acetylglucosaminyltransferase. A recombinant strain derived from GS200/LAC4 with a deletion of this gene was obtained. A study of the rate of degradation of the β-galactosidase in the deletion strain GS200/LAC4/ΔβNA showed that this strain also has impaired autophagy processes, but the rate of their passage still remains higher than the rate revealed by the insertion mutant GS200/LAC4/2.6.7. The rate of degradation of native K. phaffii enzymes formaldehyde dehydrogenase and formate dehydrogenase, which are degraded by autophagy, was also determined. Delayed degradation of native proteins was observed in both strains GS200/LAC4/2.6.7 and GS200/LAC4/ΔβNA compared to the original GS200/LAC4 strain. However, as in the case of β-galactosidase, the processes of autophagic degradation of native proteins are slower in strain GS200/LAC4/2.6.7. This may indicate the presence of other mutations in this strain that affect the process of autophagy. No genes homologous to the identified K. phaffii gene have been identified in the human genome, but genes encoding enzymes with similar function have been found. Three such genes were silenced in human cell culture HEK293, however, no changes in the processes of autophagic degradation of cytosolic proteins in human cells were observed. 1 article was published. Product Description popup.authors Zazulya Anastasiya Z. Semkiv Marta V. popup.nrat_date 2022-03-09 Close