title: Systematic identification of genes involved in metabolic acid stress resistance in yeast and their potential as cancer targets.
abstract: A hallmark of all primary and metastatic tumours is their high rate of glucose uptake and glycolysis. A consequence of the glycolytic phenotype is the accumulation of metabolic acid; hence, tumour cells experience considerable intracellular acid stress. To compensate, tumour cells upregulate acid pumps, which expel the metabolic acid into the surrounding tumour environment, resulting in alkalization of intracellular pH and acidification of the tumour microenvironment. Nevertheless, we have only a limited understanding of the consequences of altered intracellular pH on cell physiology, or of the genes and pathways that respond to metabolic acid stress. We have used yeast as a genetic model for metabolic acid stress with the rationale that the metabolic changes that occur in cancer that lead to intracellular acid stress are likely fundamental. Using a quantitative systems biology approach we identified 129 genes required for optimal growth under conditions of metabolic acid stress. We identified six highly conserved protein complexes with functions related to oxidative phosphorylation (mitochondrial respiratory chain complex III and IV), mitochondrial tRNA biosynthesis [glutamyl-tRNA(Gln) amidotransferase complex], histone methylation (Set1C-COMPASS), lysosome biogenesis (AP-3 adapter complex), and mRNA processing and P-body formation (PAN complex). We tested roles for two of these, AP-3 adapter complex and PAN deadenylase complex, in resistance to acid stress using a myeloid leukaemia-derived human cell line that we determined to be acid stress resistant. Loss of either complex inhibited growth of Hap1 cells at neutral pH and caused sensitivity to acid stress, indicating that AP-3 and PAN complexes are promising new targets in the treatment of cancer. Additionally, our data suggests that tumours may be genetically sensitized to acid stress and hence susceptible to acid stress-directed therapies, as many tumours accumulate mutations in mitochondrial respiratory chain complexes required for their proliferation.
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: John J
authors: JJ
authors: Shin
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Qurratulain
authors: Q
authors: Aftab
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Pamela
authors: P
authors: Austin
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Jennifer A
authors: JA
authors: McQueen
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Tak
authors: T
authors: Poon
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Shu Chen
authors: SC
authors: Li
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3.
authors: Barry P
authors: BP
authors: Young
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3 roskelly@mail.ubc.ca cloewen@mail.ubc.ca.
authors: Calvin D
authors: CD
authors: Roskelley
authors: Department of Cellular and Physiological Sciences, Life Sciences Institute, University of British Columbia, Vancouver, British Columbia, Canada V6T 1Z3 roskelly@mail.ubc.ca cloewen@mail.ubc.ca.
authors: Christopher J R
authors: 0000-0002-1760-5749
authors: CJ
authors: Loewen
mesh_terms: D045744:Cell Line, Tumor; D049109:Cell Proliferation; D055786:Gene Knockout Techniques; D005800:Genes, Fungal; D005820:Genetic Testing; D057809:HEK293 Cells; D006801:Humans; D006863:Hydrogen-Ion Concentration; D058990:Molecular Targeted Therapy; D009369:Neoplasms / Q000235:genetics* / Q000628:therapy*; D021122:Protein Subunits / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D013312:Stress, Physiological / Q000235:genetics*; D025262:Vacuolar Proton-Translocating ATPases / Q000378:metabolism
chemical_list: D021122:Protein Subunits; D025262:Vacuolar Proton-Translocating ATPases
keywords: AP-3 complex; Hap1 cells; Intracellular acid stress; Metabolism; Mitochondria; PAN complex