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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1995 Jan;177(2):357–363. doi: 10.1128/jb.177.2.357-363.1995

Isolation and characterization of mutants impaired in the selective degradation of peroxisomes in the yeast Hansenula polymorpha.

V I Titorenko 1, I Keizer 1, W Harder 1, M Veenhuis 1
PMCID: PMC176598  PMID: 7814324

Abstract

We have isolated a collection of peroxisome degradation-deficient (Pdd-) mutants of the yeast Hansenula polymorpha which are impaired in the selective autophagy of alcohol oxidase-containing peroxisomes. Two genes, designated PDD1 and PDD2, have been identified by complementation and linkage analyses. In both mutant strains, the glucose-induced proteolytic turnover of peroxisomes is fully prevented. The pdd1 and pdd2 mutant phenotypes were caused by recessive monogenic mutations. Mutations mapped in the PDD1 gene appeared to affect the initial step of peroxisome degradation, namely, sequestration of the organelle to be degraded by membrane multilayers. Thus, Pdd1p may be involved in the initial signalling events which determine which peroxisome will be degraded. The product of the PDD2 gene appeared to be essential for mediating the second step in selective peroxisome degradation, namely, fusion and subsequent uptake of the sequestered organelles into the vacuole. pdd1 and pdd2 mutations showed genetic interactions which suggested that the corresponding gene products may physically or functionally interact with each other.

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Selected References

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  1. Arias J. A., Moser A. B., Goldfischer S. L. Ultrastructural and cytochemical demonstration of peroxisomes in cultured fibroblasts from patients with peroxisomal deficiency disorders. J Cell Biol. 1985 May;100(5):1789–1792. doi: 10.1083/jcb.100.5.1789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Baba M., Takeshige K., Baba N., Ohsumi Y. Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization. J Cell Biol. 1994 Mar;124(6):903–913. doi: 10.1083/jcb.124.6.903. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolender R. P., Weibel E. R. A morphometric study of the removal of phenobarbital-induced membranes from hepatocytes after cessation of threatment. J Cell Biol. 1973 Mar;56(3):746–761. doi: 10.1083/jcb.56.3.746. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  5. Bystrykh L. V., de Koning W., Harder W. Dihydroxyacetone synthase from Candida boidinii KD1. Methods Enzymol. 1990;188:435–445. doi: 10.1016/0076-6879(90)88069-m. [DOI] [PubMed] [Google Scholar]
  6. Dunn W. A., Jr Studies on the mechanisms of autophagy: formation of the autophagic vacuole. J Cell Biol. 1990 Jun;110(6):1923–1933. doi: 10.1083/jcb.110.6.1923. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunn W. A., Jr Studies on the mechanisms of autophagy: maturation of the autophagic vacuole. J Cell Biol. 1990 Jun;110(6):1935–1945. doi: 10.1083/jcb.110.6.1935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Faber K. N., Swaving G. J., Faber F., Ab G., Harder W., Veenhuis M., Haima P. Chromosomal targeting of replicating plasmids in the yeast Hansenula polymorpha. J Gen Microbiol. 1992 Nov;138(11):2405–2416. doi: 10.1099/00221287-138-11-2405. [DOI] [PubMed] [Google Scholar]
  9. Goodman J. M., Trapp S. B., Hwang H., Veenhuis M. Peroxisomes induced in Candida boidinii by methanol, oleic acid and D-alanine vary in metabolic function but share common integral membrane proteins. J Cell Sci. 1990 Sep;97(Pt 1):193–204. doi: 10.1242/jcs.97.1.193. [DOI] [PubMed] [Google Scholar]
  10. Gould S. J., Keller G. A., Schneider M., Howell S. H., Garrard L. J., Goodman J. M., Distel B., Tabak H., Subramani S. Peroxisomal protein import is conserved between yeast, plants, insects and mammals. EMBO J. 1990 Jan;9(1):85–90. doi: 10.1002/j.1460-2075.1990.tb08083.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Guarente L. Synthetic enhancement in gene interaction: a genetic tool come of age. Trends Genet. 1993 Oct;9(10):362–366. doi: 10.1016/0168-9525(93)90042-g. [DOI] [PubMed] [Google Scholar]
  12. Heikoop J. C., van den Berg M., Strijland A., Weijers P. J., Just W. W., Meijer A. J., Tager J. M. Turnover of peroxisomal vesicles by autophagic proteolysis in cultured fibroblasts from Zellweger patients. Eur J Cell Biol. 1992 Apr;57(2):165–171. [PubMed] [Google Scholar]
  13. Kovács J., Réz G. Quantitative aspects of cellular autophagocytosis. Morphometric and cell fractionation studies. Revis Biol Celular. 1989;20:63–78. [PubMed] [Google Scholar]
  14. Kyhse-Andersen J. Electroblotting of multiple gels: a simple apparatus without buffer tank for rapid transfer of proteins from polyacrylamide to nitrocellulose. J Biochem Biophys Methods. 1984 Dec;10(3-4):203–209. doi: 10.1016/0165-022x(84)90040-x. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Luiken J. J., van den Berg M., Heikoop J. C., Meijer A. J. Autophagic degradation of peroxisomes in isolated rat hepatocytes. FEBS Lett. 1992 Jun 8;304(1):93–97. doi: 10.1016/0014-5793(92)80596-9. [DOI] [PubMed] [Google Scholar]
  17. Subramani S. Protein import into peroxisomes and biogenesis of the organelle. Annu Rev Cell Biol. 1993;9:445–478. doi: 10.1146/annurev.cb.09.110193.002305. [DOI] [PubMed] [Google Scholar]
  18. Titorenko V. I., Waterham H. R., Cregg J. M., Harder W., Veenhuis M. Peroxisome biogenesis in the yeast Hansenula polymorpha is controlled by a complex set of interacting gene products. Proc Natl Acad Sci U S A. 1993 Aug 15;90(16):7470–7474. doi: 10.1073/pnas.90.16.7470. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Tuttle D. L., Lewin A. S., Dunn W. A., Jr Selective autophagy of peroxisomes in methylotrophic yeasts. Eur J Cell Biol. 1993 Apr;60(2):283–290. [PubMed] [Google Scholar]
  20. Veenhuis M., Douma A., Harder W., Osumi M. Degradation and turnover of peroxisomes in the yeast Hansenula polymorpha induced by selective inactivation of peroxisomal enzymes. Arch Microbiol. 1983 Jun;134(3):193–203. doi: 10.1007/BF00407757. [DOI] [PubMed] [Google Scholar]
  21. Veenhuis M. Peroxisome biogenesis and function in Hansenula polymorpha. Cell Biochem Funct. 1992 Sep;10(3):175–184. doi: 10.1002/cbf.290100307. [DOI] [PubMed] [Google Scholar]
  22. Waterham H. R., Keizer-Gunnink I., Goodman J. M., Harder W., Veenhuis M. Development of multipurpose peroxisomes in Candida boidinii grown in oleic acid-methanol limited continuous cultures. J Bacteriol. 1992 Jun;174(12):4057–4063. doi: 10.1128/jb.174.12.4057-4063.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Yamamoto A., Masaki R., Fukui Y., Tashiro Y. Absence of cytochrome P-450 and presence of autolysosomal membrane antigens on the isolation membranes and autophagosomal membranes in rat hepatocytes. J Histochem Cytochem. 1990 Nov;38(11):1571–1581. doi: 10.1177/38.11.2212617. [DOI] [PubMed] [Google Scholar]
  24. Yamamoto A., Masaki R., Tashiro Y. Characterization of the isolation membranes and the limiting membranes of autophagosomes in rat hepatocytes by lectin cytochemistry. J Histochem Cytochem. 1990 Apr;38(4):573–580. doi: 10.1177/38.4.2319125. [DOI] [PubMed] [Google Scholar]
  25. van der Klei I. J., Harder W., Veenhuis M. Selective inactivation of alcohol oxidase in two peroxisome-deficient mutants of the yeast Hansenula polymorpha. Yeast. 1991 Nov;7(8):813–821. doi: 10.1002/yea.320070806. [DOI] [PubMed] [Google Scholar]
  26. van der Klei I. J., Veenhuis M., Nicolay K., Harder W. In vivo inactivation of peroxisomal alcohol oxidase in Hansenula polymorpha by KCN is an irreversible process. Arch Microbiol. 1989;151(1):26–33. doi: 10.1007/BF00444664. [DOI] [PubMed] [Google Scholar]

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