Does Saccharomyces cerevisiae Require Specific Post-Translational Silencing against Leaky Translation of Hac1up?

HAC1 encodes a key transcription factor that transmits the unfolded protein response (UPR) from the endoplasmic reticulum (ER) to the nucleus and regulates downstream UPR genes in Saccharomyces cerevisiae. In response to the accumulation of unfolded proteins in the ER, Ire1p oligomers splice HAC1 pre-mRNA (HAC1^u) via a non-conventional process and allow the spliced HAC1 (HAC1) to be translated efficiently. However, leaky splicing and translation of HAC1^u may occur in non-UPR cells to induce undesirable UPR. To control accidental UPR activation, multiple fail-safe mechanisms have been proposed to prevent leaky HAC1 splicing and translation and to facilitate rapid degradation of translated Hac1^up and Hac1ⁱp. Among proposed regulatory mechanisms is a degron sequence encoded at the 5' end of the HAC1 intron that silences Hac1^up expression. To investigate the necessity of an intron-encoded degron sequence that specifically targets Hac1^up for degradation, we employed publicly available transcriptomic data to quantify leaky HAC1 splicing and translation in UPR-induced and non-UPR cells. As expected, we found that HAC1 is only efficiently spliced into HAC1 and efficiently translated into Hac1ⁱp in UPR-induced cells. However, our analysis of ribosome profiling data confirmed frequent occurrence of leaky translation of HAC1 regardless of UPR induction, demonstrating the inability of translation fail-safe to completely inhibit Hac1^up production. Additionally, among 32 yeast HAC1 surveyed, the degron sequence is highly conserved by Saccharomyces yeast but is poorly conserved by all other yeast species. Nevertheless, the degron sequence is the most conserved HAC1 intron segment in yeasts. These results suggest that the degron sequence may indeed play an important role in mitigating the accumulation of Hac1^up to prevent accidental UPR activation in the Saccharomyces yeast.