ABSTRACT
Saccharomyces cerevisiae is important for protein secretion studies, yet the complexities of protein synthesis and secretion under endoplasmic reticulum (ER) stress conditions remain not fully understood. ER stress, triggered by alterations in the ER protein folding environment, poses substantial challenges to cells, especially during heterologous protein production. In this study, we used RNA-seq to analyze the transcriptional responses of yeast strains to ER stress induced by reagents such as tunicamycin (Tm) or dithiothreitol (DTT). Our gene expression analysis revealed several crucial genes, such as HMO1 and BIO5, that are involved in ER-stress tolerance. Through metabolic engineering, the best engineered strain R23 with HMO1 overexpression and BIO5 deletion, showed enhanced ER stress tolerance and improved protein folding efficiency, leading to a 2.14-fold increase in α-amylase production under Tm treatment and a 2.04-fold increase in cell density under DTT treatment. Our findings contribute to the understanding of cellular responses to ER stress and provide a basis for further investigations into the mechanisms of ER stress at the cellular level.
Subject(s)
Endoplasmic Reticulum Stress , Gene Expression Regulation, Fungal , Saccharomyces cerevisiae Proteins , Saccharomyces cerevisiae , Tunicamycin , Endoplasmic Reticulum Stress/genetics , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Saccharomyces cerevisiae Proteins/genetics , Saccharomyces cerevisiae Proteins/metabolism , Tunicamycin/pharmacology , Gene Expression Regulation, Fungal/genetics , Dithiothreitol/pharmacology , Metabolic Engineering/methods , Protein FoldingABSTRACT
Esophageal carcinoma (ESCA) is a leading cause of cancer death worldwide, despite an overall decline in the incidence of new cases. However, knowledge of gene expression signatures for risk and prognosis stratification of ESCA is inadequate. Thus, identifying novel molecular biomarkers and therapeutic targets for ESCA might improve its prognosis and treatment. The current study investigated the role of ubiquitin-specific peptidase 53 (USP53), a member of the USP family that exhibits deubiquitinating activity, in ESCA and showed that USP53 is downregulated in ESCA tissues, indicating poor prognosis. USP53 suppresses the proliferation and growth of ESCA cells in vitro and in vivo, whereas its knockdown exerts opposite effects. AMP-activated protein kinase inhibitor reverses the effects of USP53 knockdown. USP53 also inhibits glycolysis, oxidative metabolism and mitochondrial dynamics. H3K27 acetylation increases USP53 expression by binding to its promoter region. Our study reveals that USP53 is activated by H3K27 acetylation and suppresses ESCA progression by regulating cell growth and metabolism. USP53 is therefore a promising target for ESCA treatment.
