title: Inactivation of the Hansenula polymorpha PMR1 gene affects cell viability and functioning of the secretory pathway.
abstract: In yeast, functions of the endoplasmic reticulum (ER) depend on the Golgi apparatus Ca2+ pool, which is replenished by the medial-Golgi ion pump Pmr1p. Here, to dissect the role of the Golgi Ca2+ pool in protein folding and elimination of unfolded proteins in the ER, the manifestations of the pmr1 mutation in yeast Hansenula polymorpha were studied. The PMR1 gene was disrupted in a H. polymorpha diploid strain. Haploid segregants of this diploid bearing the disruption allele were viable, though they showed a severe growth defect on synthetic medium and rapidly died during storage at low temperature. Disruption of H. polymorpha PMR1 led to defects of the Golgi-hosted protein glycosylation and vacuolar protein sorting. This mutation increased the survival rate of H. polymorpha cells upon treatment with the proapoptotic drug amiodarone. Unlike Saccharomyces cerevisiae, the H. polymorpha pmr1 mutant was not hypersensitive to chemicals that induce the accumulation of unfolded proteins in the ER, indicating that the elimination of unfolded proteins from the ER was not essentially affected. At the same time, the pmr1 mutation improved the secretion of human urokinase and decreased its intracellular aggregation, indicating an influence of the mutation on the protein folding in the ER.
authors.affiliation: Institute of Experimental Cardiology, Cardiology Research Center, Moscow, Russia. moagaphonov@cardio.ru
authors.forename: M O
authors.initials: MO
authors.lastname: Agaphonov
authors.forename: T A
authors.initials: TA
authors.lastname: Plotnikova
authors.forename: A V
authors.initials: AV
authors.lastname: Fokina
authors.forename: N V
authors.initials: NV
authors.lastname: Romanova
authors.forename: A N
authors.initials: AN
authors.lastname: Packeiser
authors.forename: H A
authors.initials: HA
authors.lastname: Kang
authors.forename: M D
authors.initials: MD
authors.lastname: Ter-Avanesyan
mesh_terms: D000252:Calcium-Transporting ATPases / Q000235:genetics* / Q000502:physiology*; D005656:Fungal Proteins / Q000235:genetics / Q000502:physiology; D017353:Gene Deletion; D020043:Genes, Essential; D006031:Glycosylation; D050296:Microbial Viability / Q000235:genetics*; D016254:Mutagenesis, Insertional; D010843:Pichia / Q000235:genetics* / Q000254:growth & development / Q000502:physiology*; D021381:Protein Transport / Q000235:genetics; D011994:Recombinant Proteins / Q000378:metabolism; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator; D000252:Calcium-Transporting ATPases