Mutations affecting the expression of the MOX gene encoding peroxisomal methanol oxidase in Hansenula polymorpha

2000 ◽  
Vol 38 (4) ◽  
pp. 163-170 ◽  
Author(s):  
Veronica Vallini ◽  
Enrico Berardi ◽  
Rosanna Strabbioli
Keyword(s):  
Hansenula Polymorpha ◽  
Gene Encoding ◽  
Methanol Oxidase

Biosynthesis and regulation of the peroxisomal methanol oxidase from the methylotrophic yeast Hansenula polymorpha

1984 ◽  
Vol 194 (3) ◽  
pp. 489-493 ◽  
Author(s):  
Rainer Roggenkamp ◽  
Zbigniew Janowicz ◽  
Brigitte Stanikowski ◽  
Cornelis P. Hollenberg
Keyword(s):  
Hansenula Polymorpha ◽  
Methylotrophic Yeast ◽  
Methanol Oxidase

scholarly journals Clustering of the YNA1 gene encoding a Zn(II)2Cys6 transcriptional factor in the yeast Hansenula polymorpha with the nitrate assimilation genes YNT1, YNI1 and YNR1, and its involvement in their transcriptional activation

1998 ◽  
Vol 335 (3) ◽  
pp. 647-652 ◽  
Author(s):  
Julio ÁVILA ◽  
Celedonio GONZÁLEZ ◽  
Nélida BRITO ◽  
José M. SIVERIO
Keyword(s):  
Transcriptional Activation ◽  
Hansenula Polymorpha ◽  
Nitrate Assimilation ◽  
Transcriptional Factors ◽  
Gene Encoding ◽  
Reading Frame ◽  
Transcription Start Sites ◽  
The Family ◽  
Genes Encoding ◽  
Free Media

The genes encoding the nitrate transporter (YNT1), nitrite reductase (YNI1) and nitrate reductase (YNR1) are clustered in the yeast Hansenula polymorpha. In addition, DNA sequencing of the region containing these genes demonstrated that a new open reading frame called YNA1 (yeast nitrate assimilation) was located between YNR1 and YNI1. The YNA1 gene encodes a protein of 529 residues belonging to the family of Zn(II)2Cys6 fungal transcriptional factors, and has the highest similarity to the transcriptional factors encoded by nirA, and to a smaller extent to nit-4, involved in the nitrate induction of the gene involved in the assimilation of this compound in filamentous fungi. Northern blot analysis showed the presence of the YNA1 transcript in cells incubated in nitrate, nitrate plus ammonium, ammonium, and nitrogen-free media, with a decrease in its levels in those cells incubated in ammonium. In nitrate the strain Δyna1::URA3, with a disrupted YNA1 gene, neither grew nor expressed the genes YNT1, YNI1 and YNR1. In the gene cluster YNT1-YNI1-YNA1-YNR1, the four genes were transcribed independently in the YNT1 → YNR1 direction and the transcription start sites were determined by primer extension.

Cell wall strength of Hansenula polymorpha in continuous cultures in relation to the recovery of methanol oxidase (MOX)

1987 ◽  
Vol 27 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Marco L. F. Giuseppin ◽  
Hendrikus M. J. van Eijk ◽  
Marja Hellendoorn ◽  
José W. van Almkerk
Keyword(s):  
Cell Wall ◽  
Hansenula Polymorpha ◽  
Continuous Cultures ◽  
Wall Strength ◽  
Methanol Oxidase

Formation of irregular giant peroxisomes by overproduction of the crystalloid core protein methanol oxidase in the methylotrophic yeast Hansenula polymorpha

1989 ◽  
Vol 9 (3) ◽  
pp. 988-994
Author(s):  
R Roggenkamp ◽  
T Didion ◽  
K V Kowallik
Keyword(s):  
Protein Import ◽  
Core Protein ◽  
Hansenula Polymorpha ◽  
Selection Procedure ◽  
Methylotrophic Yeast ◽  
Cellular Protein ◽  
Yeast Cells ◽  
Thin Sections ◽  
Single Protein ◽  
Methanol Oxidase

The crystalloid core in peroxisomes of the methylotrophic yeast Hansenula polymorpha is composed of the octameric flavoprotein methanol oxidase (MOX). We transformed yeast cells with a high-copy-number vector harboring the cloned MOX gene in order to study the effects on regulation, protein import, and peroxisome biosynthesis. In transformed wild-type cells, no increase in expression of MOX was detectable. Mutants defective in MOX activity were isolated by a specific selection procedure. Two structural MOX mutants are described that allow overproduction of a fully active enzyme upon transformation at quantities of about two-thirds of the total cellular protein. The overproduced protein was imported into peroxisomes, altering their morphology (in thin sections) and stability in cell lysates; the organelles showed a tendency to form rectangular bodies, and their lumina were completely filled with the crystalloid structure. The overall size of the peroxisomes was increased severalfold in comparison with the size of nontransformed yeast cells. The results suggest high capacities of peroxisomal growth conferred by overproduction and import of a single protein.

scholarly journals Formation of irregular giant peroxisomes by overproduction of the crystalloid core protein methanol oxidase in the methylotrophic yeast Hansenula polymorpha.

1989 ◽  
Vol 9 (3) ◽  
pp. 988-994 ◽  
Author(s):  
R Roggenkamp ◽  
T Didion ◽  
K V Kowallik
Keyword(s):  
Protein Import ◽  
Core Protein ◽  
Hansenula Polymorpha ◽  
Selection Procedure ◽  
Methylotrophic Yeast ◽  
Cellular Protein ◽  
Yeast Cells ◽  
Thin Sections ◽  
Single Protein ◽  
Methanol Oxidase

The crystalloid core in peroxisomes of the methylotrophic yeast Hansenula polymorpha is composed of the octameric flavoprotein methanol oxidase (MOX). We transformed yeast cells with a high-copy-number vector harboring the cloned MOX gene in order to study the effects on regulation, protein import, and peroxisome biosynthesis. In transformed wild-type cells, no increase in expression of MOX was detectable. Mutants defective in MOX activity were isolated by a specific selection procedure. Two structural MOX mutants are described that allow overproduction of a fully active enzyme upon transformation at quantities of about two-thirds of the total cellular protein. The overproduced protein was imported into peroxisomes, altering their morphology (in thin sections) and stability in cell lysates; the organelles showed a tendency to form rectangular bodies, and their lumina were completely filled with the crystalloid structure. The overall size of the peroxisomes was increased severalfold in comparison with the size of nontransformed yeast cells. The results suggest high capacities of peroxisomal growth conferred by overproduction and import of a single protein.

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