The unfolded protein response in Pichia pastoris without external stressing stimuli.

Abstract

Dysfunction or capacity shortage of the endoplasmic reticulum (ER) is cumulatively called ER stress and provokes the unfolded protein response (UPR). In various yeast species, the ER-located transmembrane protein Ire1 is activated upon ER stress and performs the splicing reaction of HAC1 mRNA, the mature form of which is translated into a transcription factor protein that is responsible for the transcriptome change on the UPR. Here we carefully assessed the splicing of HAC1 mRNA in Pichia pastoris (Komagataella phaffii) cells. We found that, inconsistent with previous reports by others, the HAC1 mRNA was substantially, but partially, spliced even without ER-stressing stimuli. Unlike Saccharomyces cerevisiae, growth of P.pastoris was significantly retarded by the IRE1-gene knockout mutation. Moreover, P.pastoris cells seemed to push more abundant proteins into the secretory pathway than S.cerevisiae cells. We also suggest that P.pastoris Ire1 has the ability to control its activity stringently in an ER stress-dependent manner. We thus propose that P.pastoris cells are highly ER-stressed possibly because of the high load of endogenous proteins into the ER.