Vacuolar accumulation and extracellular extrusion of electrophilic compounds by wild-type and glutathione-deficient mutants of the methylotrophic yeast Hansenula polymorpha

Hansenula polymorpha ass3 mutants are characterized by the accumulation of inactive alcohol oxidase (AO) monomers in the cytosol, whereas other peroxisomal matrix proteins are normally activated and sorted to peroxisomes. These mutants also have a glutamate or aspartate requirement on minimal media. Cloning of the corresponding gene resulted in the isolation of the H. polymorpha PYC gene that encodes pyruvate carboxylase (HpPyc1p). HpPyc1p is a cytosolic, anapleurotic enzyme that replenishes the tricarboxylic acid cycle with oxaloacetate. The absence of this enzyme can be compensated by addition of aspartate or glutamate to the growth media. We show that HpPyc1p protein but not the enzyme activity is essential for import and assembly of AO. Similar results were obtained in the related yeast Pichia pastoris. In vitro studies revealed that HpPyc1p has affinity for FAD and is capable to physically interact with AO protein. These data suggest that in methylotrophic yeast pyruvate carboxylase plays a dual role in that, besides its well-characterized metabolic function as anapleurotic enzyme, the protein fulfils a specific role in the AO sorting and assembly process, possibly by mediating FAD-binding to AO monomers.

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Methanol and ethanol utilization in methylotrophic yeast Pichia pinus wild-type and mutant strains

Archives of Microbiology ◽

10.1007/bf00245392 ◽

1991 ◽

Vol 156 (6) ◽

pp. 455-462 ◽

Cited By ~ 9

Author(s):

A. A. Sibirny ◽

V. I. Titorenko ◽

G. E. Teslyar ◽

V. I. Petrushko ◽

M. M. Kucher

Keyword(s):

Methylotrophic Yeast ◽

Wild Type ◽

Mutant Strains ◽

Pichia Pinus

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Expression and targeting of a 47 kDa integral peroxisomal membrane protein of Candida boidinii in wild type and a peroxisome-deficient mutant of Hansenula polymorpha

FEBS Letters ◽

10.1016/0014-5793(93)81166-w ◽

1993 ◽

Vol 315 (3) ◽

pp. 211-216 ◽

Cited By ~ 5

Author(s):

G.J. Sulter ◽

H.R. Waterham ◽

E.G. Vrieling ◽

J.M. Goodman ◽

W. Harder ◽

...

Keyword(s):

Membrane Protein ◽

Hansenula Polymorpha ◽

Candida Boidinii ◽

Deficient Mutant ◽

Wild Type ◽

Peroxisomal Membrane ◽

Peroxisomal Membrane Protein

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Development of the Thermotolerant Methylotrophic Yeast Hansenula polymorpha as Efficient Ethanol Producer

Yeast Diversity in Human Welfare ◽

10.1007/978-981-10-2621-8_11 ◽

2017 ◽

pp. 257-282 ◽

Cited By ~ 2

Author(s):

Kostyantyn Dmytruk ◽

Olena Kurylenko ◽

Justyna Ruchala ◽

Olena Ishchuk ◽

Andriy Sibirny

Keyword(s):

Hansenula Polymorpha ◽

Methylotrophic Yeast ◽

Ethanol Producer

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Peroxisome Maintenance Depends on De Novo Peroxisome Formation in Yeast Mutants Defective in Peroxisome Fission and Inheritance

International Journal of Molecular Sciences ◽

10.3390/ijms20164023 ◽

2019 ◽

Vol 20 (16) ◽

pp. 4023 ◽

Cited By ~ 2

Author(s):

Justyna P. Wróblewska ◽

Ida J. van der Klei

Keyword(s):

De Novo ◽

Hansenula Polymorpha ◽

Yeast Cells ◽

Wild Type ◽

Double Deletion ◽

Mother Cells ◽

Yeast Mutants ◽

Rescue Mechanism ◽

Wild Type Yeast ◽

In Cells

There is an ongoing debate on how peroxisomes form: by growth and fission of pre-existing peroxisomes or de novo from another membrane. It has been proposed that, in wild type yeast cells, peroxisome fission and careful segregation of the organelles over mother cells and buds is essential for organelle maintenance. Using live cell imaging we observed that cells of the yeast Hansenula polymorpha, lacking the peroxisome fission protein Pex11, still show peroxisome fission and inheritance. Also, in cells of mutants without the peroxisome inheritance protein Inp2 peroxisome segregation can still occur. In contrast, peroxisome fission and inheritance were not observed in cells of a pex11 inp2 double deletion strain. In buds of cells of this double mutant, new organelles likely appear de novo. Growth of pex11 inp2 cells on methanol, a growth substrate that requires functional peroxisomes, is retarded relative to the wild type control. Based on these observations we conclude that in H. polymorpha de novo peroxisome formation is a rescue mechanism, which is less efficient than organelle fission and inheritance to maintain functional peroxisomes.

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