title: A hypomorphic allele of the first N-glycosylation gene, ALG7, causes mitochondrial defects in yeast.
abstract: The modification of proteins at asparagine residues with oligosaccharides (N-glycans) plays critical roles in diverse cell functions. N-glycans originate from a common lipid-linked oligosaccharide (LLO) precursor whose synthesis is initiated by the Dol-P-dependent GlcNAc-1-P transferase (GPT) encoded by an essential ALG7 gene. To identify cellular processes affected by ALG7 and N-glycosylation, we replaced the genomic copy of ALG7 with its hypomorphic allele in two genetically distinct haploid yeast cells. We show that ALG7 knockdown gave rise to an unexpected phenotype of mitochondrial dysfunction. The alg7 mutants did not grow on glycerol and DNA arrays revealed the absence of mitochondrial genes' expression. Accordingly, the alg7 mutants displayed no detectable mtDNA and respiratory activity. Both mutants exhibited diminished abundance of LLO and under-glycosylation of carboxypeptidase Y (CPY). Moreover, another N-glycosylation mutant with a LLO defect, alg6, was respiratory deficient. Collectively, our studies provide evidence that the dysregulation of N-glycosylation in haploid yeast cells leads to mitochondrial dysfunction.
authors.affiliation: Department of Biochemistry, School of Medicine, Boston University Medical Center, Boston, MA, United States.
authors.forename: Richard D
authors.initials: RD
authors.lastname: Mendelsohn
authors.forename: Eva J
authors.initials: EJ
authors.lastname: Helmerhorst
authors.forename: John F
authors.initials: JF
authors.lastname: Cipollo
authors.forename: Maria A
authors.initials: MA
authors.lastname: Kukuruzinska
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000483:Alleles; D004272:DNA, Mitochondrial / Q000032:analysis; D006031:Glycosylation; D008928:Mitochondria / Q000502:physiology*; D010101:Oxygen Consumption; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000235:genetics*
chemical_list: D020413:3' Untranslated Regions; D004272:DNA, Mitochondrial; D017855:Transferases (Other Substituted Phosphate Groups); C023222:UDPacetylglucosamine-dolichyl-phosphate acetylglucosamine-1-phosphate transferase