Limits...
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.

Show MeSH

Related in: MedlinePlus

The H. polymorpha Pex11 family members. Fluorescence microscopy images of methanol-grown WT cells producing Pex11-GFP (A), Pex25-GFP (B), or Pex11C-GFP (C). All three proteins are localized to peroxisomes. (D–F) Fluorescence microscopy images of pex11 pex25 (D), pex11 pex11C (E), and pex25 pex11C cells (F) producing DsRed-SKL to mark the peroxisomal matrix. Cells were grown on glycerol/methanol mixtures. The DsRed-SKL fluorescence does not completely fill the matrix of the peroxisomes because of the presence of alcohol oxidase crystal inside the peroxisomes. Bar, 1 µm. Images were taken by wide-field fluorescence microscopy. The cell contour is indicated in blue. (G) Western blots showing the levels of Pex3 and Pex14 proteins in WT and various deletion strains. Cells were grown for 16 h on glycerol/methanol. Equal amounts of protein were loaded per lane. The first lane shows the negative controls of the corresponding deletion strain (i.e., pex3 and pex14). The pyruvate carboxylase (Pyc1) blot is added as loading control.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC3105547&req=5

fig1: The H. polymorpha Pex11 family members. Fluorescence microscopy images of methanol-grown WT cells producing Pex11-GFP (A), Pex25-GFP (B), or Pex11C-GFP (C). All three proteins are localized to peroxisomes. (D–F) Fluorescence microscopy images of pex11 pex25 (D), pex11 pex11C (E), and pex25 pex11C cells (F) producing DsRed-SKL to mark the peroxisomal matrix. Cells were grown on glycerol/methanol mixtures. The DsRed-SKL fluorescence does not completely fill the matrix of the peroxisomes because of the presence of alcohol oxidase crystal inside the peroxisomes. Bar, 1 µm. Images were taken by wide-field fluorescence microscopy. The cell contour is indicated in blue. (G) Western blots showing the levels of Pex3 and Pex14 proteins in WT and various deletion strains. Cells were grown for 16 h on glycerol/methanol. Equal amounts of protein were loaded per lane. The first lane shows the negative controls of the corresponding deletion strain (i.e., pex3 and pex14). The pyruvate carboxylase (Pyc1) blot is added as loading control.

Mentions: As shown in Fig. 1, A–C, all three members of the Pex11 protein family are localized to peroxisomes. The fluorescence signal observed for Pex11C-GFP is low relative to Pex11-GFP and Pex25-GFP, which is most likely due to relatively low expression of PEX11C as also is suggested by transcriptomics data (van Zutphen et al., 2010).


Peroxisome reintroduction in Hansenula polymorpha requires Pex25 and Rho1.

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

The H. polymorpha Pex11 family members. Fluorescence microscopy images of methanol-grown WT cells producing Pex11-GFP (A), Pex25-GFP (B), or Pex11C-GFP (C). All three proteins are localized to peroxisomes. (D–F) Fluorescence microscopy images of pex11 pex25 (D), pex11 pex11C (E), and pex25 pex11C cells (F) producing DsRed-SKL to mark the peroxisomal matrix. Cells were grown on glycerol/methanol mixtures. The DsRed-SKL fluorescence does not completely fill the matrix of the peroxisomes because of the presence of alcohol oxidase crystal inside the peroxisomes. Bar, 1 µm. Images were taken by wide-field fluorescence microscopy. The cell contour is indicated in blue. (G) Western blots showing the levels of Pex3 and Pex14 proteins in WT and various deletion strains. Cells were grown for 16 h on glycerol/methanol. Equal amounts of protein were loaded per lane. The first lane shows the negative controls of the corresponding deletion strain (i.e., pex3 and pex14). The pyruvate carboxylase (Pyc1) blot is added as loading control.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC3105547&req=5

fig1: The H. polymorpha Pex11 family members. Fluorescence microscopy images of methanol-grown WT cells producing Pex11-GFP (A), Pex25-GFP (B), or Pex11C-GFP (C). All three proteins are localized to peroxisomes. (D–F) Fluorescence microscopy images of pex11 pex25 (D), pex11 pex11C (E), and pex25 pex11C cells (F) producing DsRed-SKL to mark the peroxisomal matrix. Cells were grown on glycerol/methanol mixtures. The DsRed-SKL fluorescence does not completely fill the matrix of the peroxisomes because of the presence of alcohol oxidase crystal inside the peroxisomes. Bar, 1 µm. Images were taken by wide-field fluorescence microscopy. The cell contour is indicated in blue. (G) Western blots showing the levels of Pex3 and Pex14 proteins in WT and various deletion strains. Cells were grown for 16 h on glycerol/methanol. Equal amounts of protein were loaded per lane. The first lane shows the negative controls of the corresponding deletion strain (i.e., pex3 and pex14). The pyruvate carboxylase (Pyc1) blot is added as loading control.
Mentions: As shown in Fig. 1, A–C, all three members of the Pex11 protein family are localized to peroxisomes. The fluorescence signal observed for Pex11C-GFP is low relative to Pex11-GFP and Pex25-GFP, which is most likely due to relatively low expression of PEX11C as also is suggested by transcriptomics data (van Zutphen et al., 2010).

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