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Peroxisome reintroduction in Hansenula polymorpha requires Pex25 and Rho1.

Saraya R, Krikken AM, Veenhuis M, van der Klei IJ - J. Cell Biol. (2011)

Bottom Line: Pex25 cells were not themselves peroxisome deficient but instead contained a slightly increased number of peroxisomes.Peroxisomes reappeared in pex11 pex25 cells upon synthesis of Pex25, but not of Pex11.These data therefore provide new and detailed insight into factors important for de novo peroxisome formation in yeast.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute, Kluyver Centre for Genomics of Industrial Fermentation, University of Groningen, 9700 CC Groningen, Netherlands.

ABSTRACT
We identified two proteins, Pex25 and Rho1, which are involved in reintroduction of peroxisomes in peroxisome-deficient yeast cells. These are, together with Pex3, the first proteins identified as essential for this process. Of the three members of the Hansenula polymorpha Pex11 protein family-Pex11, Pex25, and Pex11C-only Pex25 was required for reintroduction of peroxisomes into a peroxisome-deficient mutant strain. In peroxisome-deficient pex3 cells, Pex25 localized to structures adjacent to the ER, whereas in wild-type cells it localized to peroxisomes. Pex25 cells were not themselves peroxisome deficient but instead contained a slightly increased number of peroxisomes. Interestingly, pex11 pex25 double deletion cells, in which both peroxisome fission (due to the deletion of PEX11) and reintroduction (due to deletion of PEX25) was blocked, did display a peroxisome-deficient phenotype. Peroxisomes reappeared in pex11 pex25 cells upon synthesis of Pex25, but not of Pex11. Reintroduction in the presence of Pex25 required the function of the GTPase Rho1. These data therefore provide new and detailed insight into factors important for de novo peroxisome formation in yeast.

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Peroxisomes are formed in pex11 pex25 cells upon reintroduction of PEX25, but not upon reintroduction of PEX11. (A–C) pex11 pex25 PAOX PEX25-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol-containing media. Pex25-mCherry fluorescence is shown in the images in the left panels (in red). Cells were grown for 4 (A and B) or 16 h (C). (D–F) pex11 pex25 PAOX PEX11-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol medium. Cells were grown for 4 (D and E) or 16 h (F). The images at the right show the merged fluorescence images as well as the cell walls in blue. Bar, 1 µm.
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fig6: Peroxisomes are formed in pex11 pex25 cells upon reintroduction of PEX25, but not upon reintroduction of PEX11. (A–C) pex11 pex25 PAOX PEX25-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol-containing media. Pex25-mCherry fluorescence is shown in the images in the left panels (in red). Cells were grown for 4 (A and B) or 16 h (C). (D–F) pex11 pex25 PAOX PEX11-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol medium. Cells were grown for 4 (D and E) or 16 h (F). The images at the right show the merged fluorescence images as well as the cell walls in blue. Bar, 1 µm.

Mentions: We next analyzed whether peroxisomes were formed in pex11 pex25 cells upon reintroduction of either PEX25-mCherry (Fig. 6, A–C) or PEX11-mCherry (Fig. 6, D–F). Separate strains were constructed in which either the ER marker protein BiPN30-GFP-HDEL (Fig. 6, A and D) or Pex3-GFP (Fig. 6, B and E) or GFP-SKL (Fig. 6, C and F) were produced. Upon a shift of pex11 pex25 PAOX PEX25-mCherry cells from PAOX-repressing to PAOX-inducing conditions (shift from glucose/ammonium sulfate to methanol/glycerol/ammonium sulfate), peroxisomes were readily formed (Fig. 6 C). After 4 h of incubation in the presence of methanol, Pex25 was initially localized in a spot at the ER (Fig. 6 A). Pex3 colocalized with Pex25 at this spot (Fig. 6 B). At later time points (16 h after induction) the cells grew on methanol like pex11 cells and contained peroxisomes harboring the peroxisomal matrix marker protein GFP-SKL (Fig. 6 C).


Peroxisome reintroduction in Hansenula polymorpha requires Pex25 and Rho1.

Saraya R, Krikken AM, Veenhuis M, van der Klei IJ - J. Cell Biol. (2011)

Peroxisomes are formed in pex11 pex25 cells upon reintroduction of PEX25, but not upon reintroduction of PEX11. (A–C) pex11 pex25 PAOX PEX25-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol-containing media. Pex25-mCherry fluorescence is shown in the images in the left panels (in red). Cells were grown for 4 (A and B) or 16 h (C). (D–F) pex11 pex25 PAOX PEX11-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol medium. Cells were grown for 4 (D and E) or 16 h (F). The images at the right show the merged fluorescence images as well as the cell walls in blue. Bar, 1 µm.
© Copyright Policy - openaccess
Related In: Results  -  Collection

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fig6: Peroxisomes are formed in pex11 pex25 cells upon reintroduction of PEX25, but not upon reintroduction of PEX11. (A–C) pex11 pex25 PAOX PEX25-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol-containing media. Pex25-mCherry fluorescence is shown in the images in the left panels (in red). Cells were grown for 4 (A and B) or 16 h (C). (D–F) pex11 pex25 PAOX PEX11-mCherry cells were shifted from glucose/ammonium sulfate to glycerol/methanol medium. Cells were grown for 4 (D and E) or 16 h (F). The images at the right show the merged fluorescence images as well as the cell walls in blue. Bar, 1 µm.
Mentions: We next analyzed whether peroxisomes were formed in pex11 pex25 cells upon reintroduction of either PEX25-mCherry (Fig. 6, A–C) or PEX11-mCherry (Fig. 6, D–F). Separate strains were constructed in which either the ER marker protein BiPN30-GFP-HDEL (Fig. 6, A and D) or Pex3-GFP (Fig. 6, B and E) or GFP-SKL (Fig. 6, C and F) were produced. Upon a shift of pex11 pex25 PAOX PEX25-mCherry cells from PAOX-repressing to PAOX-inducing conditions (shift from glucose/ammonium sulfate to methanol/glycerol/ammonium sulfate), peroxisomes were readily formed (Fig. 6 C). After 4 h of incubation in the presence of methanol, Pex25 was initially localized in a spot at the ER (Fig. 6 A). Pex3 colocalized with Pex25 at this spot (Fig. 6 B). At later time points (16 h after induction) the cells grew on methanol like pex11 cells and contained peroxisomes harboring the peroxisomal matrix marker protein GFP-SKL (Fig. 6 C).

Bottom Line: Pex25 cells were not themselves peroxisome deficient but instead contained a slightly increased number of peroxisomes.Peroxisomes reappeared in pex11 pex25 cells upon synthesis of Pex25, but not of Pex11.These data therefore provide new and detailed insight into factors important for de novo peroxisome formation in yeast.

View Article: PubMed Central - HTML - PubMed

Affiliation: Molecular Cell Biology, Groningen Biomolecular Sciences and Biotechnology Institute, Kluyver Centre for Genomics of Industrial Fermentation, University of Groningen, 9700 CC Groningen, Netherlands.

ABSTRACT
We identified two proteins, Pex25 and Rho1, which are involved in reintroduction of peroxisomes in peroxisome-deficient yeast cells. These are, together with Pex3, the first proteins identified as essential for this process. Of the three members of the Hansenula polymorpha Pex11 protein family-Pex11, Pex25, and Pex11C-only Pex25 was required for reintroduction of peroxisomes into a peroxisome-deficient mutant strain. In peroxisome-deficient pex3 cells, Pex25 localized to structures adjacent to the ER, whereas in wild-type cells it localized to peroxisomes. Pex25 cells were not themselves peroxisome deficient but instead contained a slightly increased number of peroxisomes. Interestingly, pex11 pex25 double deletion cells, in which both peroxisome fission (due to the deletion of PEX11) and reintroduction (due to deletion of PEX25) was blocked, did display a peroxisome-deficient phenotype. Peroxisomes reappeared in pex11 pex25 cells upon synthesis of Pex25, but not of Pex11. Reintroduction in the presence of Pex25 required the function of the GTPase Rho1. These data therefore provide new and detailed insight into factors important for de novo peroxisome formation in yeast.

Show MeSH
Related in: MedlinePlus