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Degradation and turnover of peroxisomes in the yeast Hansenula polymorpha induced by selective inactivation of peroxisomal enzymes

Abstract

Inactivation of peroxisomal enzymes in the yeast Hansenula polymorpha was studied following transfer of cells into cultivation media in which their activity was no longer required for growth. After transfer of methanol-grown cells into media containing glucose — a substrate that fully represses alcohol oxidase synthesis — the rapid inactivation of alcohol oxidase and catalase was paralleled by a disappearance of alcohol oxidase and catalase protein. The rate and extent of this inactivation was dependent upon conditions of cultivation of cells prior to their transfer. This carbon catabolite inactivation of alcohol oxidase was paralleled by degradation of peroxisomes which occurred by means of an autophagic process that was initiated by the formation of a number of electron-dense membranes around the organelles to be degraded. Sequestration was confined to peroxisomes; other cell-components such as ribosomes were absent in the sequestered cell compartment. Also, cytochemically, hydrolytic enzymes could not be demonstrated in these autophagosomes. The vacuole played a major role in the subsequent peroxisomal breakdown since it provided the enzymes required for proteolysis. Two basically similar mechanisms were observed with respect to the administration of vacuolar enzymes into the sequestered cell compartment. The first mechanism involved incorporation of a small vacuolar vesicle into the sequestered cell compartment. The delimiting membrane of this vacuolar vesicle subsequently disrupted, thereby exposing the contents of the sequestered cell compartment to vacuolar hydrolases which then degraded the peroxisomal proteins. The second mechanism, observed in cells which already contained one or more autophagic vacuoles, included fusion of the delimiting membranes of an autophagosome with the membrane surrounding an autophagic vacuole which led to migration of the peroxisome inside the latter organelle. Peroxisomes of methanolgrown H. polymorpha were degraded individually. In one cell 2 or 3 peroxisomes might be subject to degradation at the same time, but they were never observed together in one autophagosome. However, fusions of autophagic vacuoles in one cell were frequently observed. After inhibition of the cell's energy-metabolism by cyanide ions or during anaerobic incubations the formation of autophagosomes was prevented and degradation was not observed.

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References

  1. Betz H, Weiser U (1976a) Protein degradation and proteinases during yeast sporulation. Eur J Biochem 62:65–76

  2. Betz H, Weiser U (1976b) Protein degradation during yeast sporulation. Enzyme and cytochrome patterns. Eur J Biochem 70:385–395

  3. Bormann C (1980) Untersuchungen zum Abbau der peroxisomalen Alkohol-Oxidase in Candida boidinii. PhD Thesis, University of Düsseldorf, FRG

  4. Bormann C, Sahm H (1978) Degradation of microbodies in relation to activities of alcohol oxidase and catalase in Candida boidinii. Arch Microbiol 117:67–72

  5. Bruinenberg PG, Veenhuis M, Dijken JP van, Duine JA, Harder W (1982) A quantitative analysis of selective inactivation of peroxisomal enzymes in the yeast Hansenula polymorpha by high-performance liquid chromatography. FEMS Microbiol Lett 15:45–50

  6. Dijken JP van (1976) Oxidation of methanol by yeasts. PhD Thesis, University of Groningen, The Netherlands

  7. Dijken JP van, Otto R, Harder W (1976) Growth of Hansenula polymorpha in a methanol-limited chemostat. Physiological responses due to the involvement of methanol oxidase as a key enzyme in methanol metabolism. Arch Microbiol 111:137–144

  8. Eggeling L, Sahm H (1978) Derepression and partial insensitivity to carbon catabolite repression of methanol dissimilating enzymes in Hansenula polymorpha. Europ J Appl Microbiol Biotechnol 5:197–202

  9. Egli Th (1980) Wachstum von Methanol assimilierenden Hefen. PhD Thesis, Eidgenössische Technische Hochschule Zürich, Switzerland

  10. Egli Th, Dijken JP van, Veenhuis M, Harder W, Fiechter A (1980) Methanol metabolism in yeasts: regulation of the synthesis of catabolic enzymes. Arch Microbiol 124:115–121

  11. Gancedo C (1971) Inactivation of fructose-1,6-diphosphatase in glucose in yeast. J Bacteriol 107:401–405

  12. Glaumann H, Ericsson JLE, Marzella L (1980) Mechanisms of intralysosomal degradation with special reference to autophagocytosis and heterophagocytosis of cell organelles. Int Rev Cytobiol 73:149–182

  13. Gordon CN (1972) The use of octadecanol monolayers as wetting agents in the negative staining technique. J Ultrastruct Res 39:173–185

  14. Hemmings BA (1978) Evidence for the degradation of nicotinamide adenine dinucleotide phosphate-dependent glutamate dehydrogenase of Candida utilis during rapid enzyme inactivation. J Bacteriol 133:867–877

  15. Holtzman E (1976) Lysosomes: a survey. Cell biology monographs: Continuation of protoplasmatologica, vol 3. Springer, Wien New York

  16. Holzer H (1976) Catabolite inactivation in yeast. TIBS 1:178–181

  17. Kitamura K, Kaneda T, Yamamoto Y (1971) Lysis of viable cells by enzymes of Arthrobacter luteus. Arch Biochem Biophys 145: 402–404

  18. Locke M, McMahon JT (1971) The origin and fate of microbodies in the fat body of an insect. J Cell Biol 48:61–78

  19. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193: 265–275

  20. Lück H (1963) Catalase. In: Bergmeyer HU (ed) Methods of enzymatic analysis. Academic Press, New York London, pp 885–894

  21. Matile Ph (1975) The lytic compartment of cells. Cell biology monographs: Continuation of protoplasmatologia, vol I. Springer, Wien New York

  22. Mazón MJ (1978) Effect of glucose starvation on the nicotin adenin denucleotide phosphate-dependent glutamate dehydrogenase of yeast. J Bacteriol 133:780–785

  23. Müller D, Holzer H (1981) Regulation of fructose-1,6-bisphosphatase in yeast by phosphorylation/dephosphorylation. Biochem Biophys Res Commun 103:926–933

  24. Neeff J, Hägele E, Neuhaus J, Heer U, Mecke D (1978) Evidence for catabolite degradation in the glucose-dependent inactivation of yeast cytoplasmic malate dehydrogenase. Eur J Biochem 87: 489–495

  25. Osumi M, Imaizumi F, Imai M, Sato H, Yamaguchi H (1975) Isolation and characterization of microbodies from Candida tropicalis PK233 cells grown on normal alkanes. J Gen Appl Microbiol 21:375–387

  26. Switzer RL (1977) The inactivation of microbial enzymes in vivo. Ann Rev Microbiol 31:135–157

  27. Tokuyashu KT (1978) A study of positive staining of ultrathin frozen sections. J Ultrastruct Res 63:287–307

  28. Veenhuis M, Dijken JP van, Harder W (1976) Cytochemical studies on the localization of methanol oxidase and other oxidases in peroxisomes of methanol-grown Hansenula polymorpha. Arch Microbiol 111:123–135

  29. Veenhuis M, Dijken JP van, Pilon SAF, Harder W (1978a) Development of crystalline peroxisomes in methanol-grown cells of the yeast Hansenula polymorpha and its relation to environmental conditions. Arch Microbiol 117:153–163

  30. Veenhuis M, Zwart K, Harder W (1978b) Degradation of peroxisomes after transfer of methanol-grown Hansenula polymorpha into glucose-containing media. FEMS Microbiol Lett 4:283–286

  31. Veenhuis M, Keizer I, Harder W (1979) Characterization of peroxisomes in glucose-grown Hansenula polymorpha and their development after the transfer of cells into methanol-containing media. Arch Microbiol 120:165–175

  32. Veenhuis M, Dijken JP van, Harder W (1980a) A new method for the cytochemical demonstration of phosphatase activities in yeasts based on the use of cerous ions. FEMS Microbiol Lett 9:285–291

  33. Veenhuis M, Dijken JP van, Harder W (1980b) In vivo inactivation of alcohol oxidase (EC 1.2.3.1) in the yeast Hansenula polymorpha. In: Bredero P, Priester W de (eds) Proceedings of the 7th European Congress on Electron Microscopy, vol II. Seventh EUREM Foundation. Leiden, The Netherlands, pp 84–85

  34. Veenhuis M, Zwart KB, Harder W (1981a) Biogenesis and turnover of peroxisomes involved in the concurrent oxidation of methanol and methylamine in Hansenula polymorpha. Arch Microbiol 129:35–41

  35. Veenhuis M, Dijken JP van, Harder W, Mayer F (1981b) Substructure of crystalline peroxisomes in methanol-grown Hansenula polymorpha: evidence for an in vivo crystal of alcohol oxidase. Molec Cell Biol 1:949–957

  36. Veenhuis M, Dijken JP van, Harder W (1983) The significance of peroxisomes in the metabolism of one-carbon compounds in yeasts. Adv Micr Physiol 24

  37. Wiemken A, Schellenberg M, Urech K (1979) Vacuoles: the sole compartment of digestive enzymes in yeast (Saccharomyces cerevisiae)? Arch Microbiol 123:23–35

  38. Wolf DH, Holzer H (1978) Proteolysis in yeast. In: Paine JV (ed) Transport and utilization of amino acids, peptides and proteins by microorganisms. John Wiley & Sons, Chichester, England

  39. Zwart KB (1983) Metabolic significance of microbodies in the yeasts Candida utilis and Hansenula polymorpha. PhD Thesis, University of Groningen, The Netherlands

  40. Zwart KB, Veenhuis M, Dijken JP van, Harder W (1980) Development of amine oxidase containing peroxisomes in yeasts during growth on glucose in the presence of methylamine as the sole source of nitrogen. Arch Microbiol 126:117–126

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Correspondence to Wim Harder.

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Veenhuis, M., Douma, A., Harder, W. et al. Degradation and turnover of peroxisomes in the yeast Hansenula polymorpha induced by selective inactivation of peroxisomal enzymes. Arch. Microbiol. 134, 193–203 (1983). https://doi.org/10.1007/BF00407757

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Key words

  • Peroxisome
  • Degradation
  • Autophagy
  • Catabolite inactivation
  • Alcohol oxidase
  • Catalase
  • Cytochemical staining
  • Ultracryotomy
  • Hansenula polymorpha