PJB-2022-315
Cellular response to the regulation of HSP70B in Chlamydomonas Reinhardtii under hight light
Tuba Sevgi
Abstract
Chlamydomonas reinhardtii is used since decades as a model organism for studying several aspects of cell biology. In this study a vector system was used for the inducible downregulation of HSP70B using a micro-RNA approach. This system is based on the nitrate reductase (NIT1) promoter, which is repressed in cells grown on ammonium and induced in cells grown on nitrate as nitrogen source. HSP70B is involved in folding of nascent peptides and refolding of denatured proteins, for instance which accumulate in high concentrations during stress periods. In this study wild type (WT) and HSP70B-knock-down strain (amiHSP70B mutant) of C. reinhardtii were investigated by the means of the kinetics of various cellular responses to the inducible downregulation of HSP70B. Cells grown under mixotrophic conditions at high light intensities. These responses was monitored at the levels of cell numbers and cell size, protein and chlorophyll concentrations, the abundance of selected target genes and proteins, and protein complex composition. As a result of the study the cell concentration was decreased although, the cell size was increased in amiHSP70B mutant. It was found that HSP70B and VIPP1 mRNA levels were significantly downregulated in mutant. The protein levels of other detected proteins (CDJ4, SecA, VIPP1) in mutant were decreased at the end of 24h. With the native-page it was approved that CDJ4 and CGE1 form complexes in the cell
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