The relevance of the phosphatidylinositolphosphat-binding motif FRRGT of Atg18 and Atg21 for the Cvt pathway and autophagy

FEBS Lett. 2006 Aug 21;580(19):4632-8. doi: 10.1016/j.febslet.2006.07.041. Epub 2006 Jul 21.

Abstract

Atg18 and Atg21 are homologous S. cerevisiae autophagy proteins. Atg18 is essential for biogenesis of Cvt vesicles and autophagosomes, while Atg21 is only essential for Cvt vesicle formation. We found that mutated Atg18-(FTTGT), which lost almost completely its binding to PtdIns3P and PtdIns(3,5)P(2), is non-functional during the Cvt pathway but active during autophagy and pexophagy. Since the Cvt pathway does not depend on PtdIns(3,5)P(2), we conclude that the Cvt pathway requires binding of Atg18 to PtdIns3P. Mutated Atg21-(FTTGT) is inactive during the Cvt pathway but showed only partly reduced binding to PtdIns-phosphates, suggesting further lipid binding domains in Atg21. GFP-Atg18-(FTTGT) and Atg21-(FTTGT)-GFP are released from vacuolar punctae to the cytosol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Motifs*
  • Autophagy*
  • Autophagy-Related Proteins
  • Base Sequence
  • Binding Sites
  • DNA Primers
  • Endopeptidases / chemistry
  • Endopeptidases / genetics
  • Endopeptidases / metabolism*
  • Lipid Metabolism
  • Membrane Proteins
  • Mutagenesis, Site-Directed
  • Phosphatidylinositol Phosphates / metabolism*
  • Protein Binding
  • Saccharomyces cerevisiae / metabolism*
  • Saccharomyces cerevisiae / physiology
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / genetics
  • Saccharomyces cerevisiae Proteins / metabolism*

Substances

  • ATG18 protein, S cerevisiae
  • Autophagy-Related Proteins
  • DNA Primers
  • Membrane Proteins
  • Phosphatidylinositol Phosphates
  • Saccharomyces cerevisiae Proteins
  • ATG21 protein, S cerevisiae
  • Endopeptidases