Characterization of the Hansenula polymorpha CPY gene encoding carboxypeptidase Y

AR Bellu, IJ Van der Klei, KB Rechinger, M Veenhuis, JAKW Kiel*

*Corresponding author for this work

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Abstract

We have isolated the Hansenula polymorpha CPY gene encoding carboxypeptidase Y (Hp-CPY). The deduced amino acid sequence revealed that Hp-CPY consists of 541 amino acids and has a calculated Mr of 60,793. The protein is highly similar to Saccharomyces cerevisiae CPY (61.8% identity). At the N-terminus of Hp-CPY signals for the entry into the secretory pathway and subsequent sorting to the vacuole were identified. Immunocytochemically, using monospecific antibodies raised against Hp-CPY, the protein was localized to the vacuole. On Western blots, a diffuse protein band was observed in extracts of H. polymorpha cells, suggesting that the protein is glycosylated. This was confirmed by endoglycosidase H treatment, which resulted in a strong reduction of the apparent Mr of the protein. We have investigated the effect of CPP deletion on the degradation of peroxisomes, an autophagous process that occurs when the organelles become redundant for growth. In Delta cpy cells peroxisomal proteins were degraded in the vacuole as efficiently as in wild-type H. polymorpha cells, indicating that CPY is not a major proteinase in this pathway. Copyright (C) 1999 John Wiley & Sons, Ltd.

Original languageEnglish
Pages (from-to)181-189
Number of pages9
JournalYeast
Volume15
Issue number3
DOIs
Publication statusPublished - Feb-1999

Keywords

  • autophagy
  • glycosylation
  • peroxisome turnover
  • methylotrophic yeast
  • vacuolar proteinases
  • SELECTIVE INACTIVATION
  • PEROXISOMAL ENZYMES
  • PICHIA-PASTORIS
  • YEAST
  • PROPEPTIDE
  • DEGRADATION
  • BIOGENESIS
  • PROTEIN
  • CLONING

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