Identification of an Exceptionally Long Intron in theHAC1Gene ofCandida parapsilosis
ABSTRACTThe unfolded protein response (UPR) in the endoplasmic reticulum (ER) is well conserved in eukaryotes from metazoa to yeast. The transcription factorHAC1is a major regulator of the UPR in many eukaryotes. DeletingHAC1in the yeastCandida parapsilosisrendered cells more sensitive to DTT, a known inducer of the UPR. The deletion strain was also sensitive to Congo red, calcofluor white, and the antifungal drug ketoconazole, indicating thatHAC1has a role in cell wall maintenance. Transcriptomic analysis revealed that treatment of the wild type with DTT resulted in the increased expression of 368 genes. Comparison with mutant cells treated with DTT reveals that expression of 137 of these genes requiresHAC1. Enriched GO term analysis includes response to ER stress, cell wall biogenesis and glycosylation. Orthologs of many of these are associated with UPR inSaccharomyces cerevisiaeandCandida albicans. Unconventional splicing of an intron fromHAC1mRNA is required to produce a functional transcription factor. The spliced intron varies in length from 19 bases inC. albicansto 379 bases inCandida glabrata, but has not been previously identified inCandida parapsilosisand related species. We used RNA-seq data andin silicoanalysis to identify theHAC1intron in 12 species in the CTG-Ser1 clade. We show that the intron has undergone major contractions and expansions in this clade, reaching up to 848 bases. Exposure to DTT induced splicing of the long intron inC. parapsilosisHAC1, inducing the UPR.IMPORTANCEThe unfolded protein response (UPR) responds to the build-up of misfolded proteins in the endoplasmic reticulum. The UPR has wide-ranging functions from fungal pathogenesis to applications in biotechnology. The UPR is regulated through the splicing of an unconventional intron in theHAC1gene. This intron has been described in many fungal species and is of variable length. Until now it was believed that some members of the CTG-Ser1 clade such asC. parapsilosisdid not contain an intron inHAC1, suggesting that the UPR was regulated in a different manner. Here we demonstrate thatHAC1plays an important role in regulating the UPR inC. parapsilosis. We also identified an unusually long intron (626 bp) inC. parapsilosisHAC1. Further analysis showed thatHAC1orthologs in several species in the CTG-Ser1 clade contain long introns.