Functional characterization of the unconventional splicing of Yarrowia lipolytica HAC1 mRNA induced by unfolded protein response

Abstract In the yeast Saccharomyces cerevisiae , the unfolded protein response (UPR) involves the unconventional splicing of HAC1 mRNA, which is mediated by the activated Ire1p transmembrane kinase/endonuclease. In this study, we isolated and characterized a Yarrowia lipolytica HAC1 ( YlHAC1 ) encoding a basic‐leucine zipper transcription factor. The null mutant strain of YlHAC1 (Δ Ylhac1 ) displayed a significantly increased sensitivity to dithiothreitol (DTT) and tunicamycin (TM), along with a defect in hyphal growth, suggesting the essential function of YlHAC1 in UPR. The unconventional splicing of YlHAC1 mRNA occurred under the UPR conditions induced by DTT or TM treatment. Unlike S. cerevisiae HAC1 mRNA with an intron of 252 nt, YlHAC1 mRNA was shown to harbour a short intron of length 29 nt. The YlHAC1 mRNA harboured the nucleotides CAG, conserved at the intron borders in the filamentous fungi hac1/hacA and mammalian XBP1 , as well as a conserved bipartite element within the 3′ untranslated region. The expression of the spliced form of YlHAC1 mRNA in the wild‐type andΔ Ylhac1 strains resulted in an increased resistance to DTT, thereby indicating that the spliced form is translated into a functional YlHac1p. Copyright © 2010 John Wiley & Sons, Ltd.