Biochemical and Biophysical Research Communications
Characterization of a novel autophagy-specific gene, ATG29☆
Section snippets
Materials and methods
Yeast strains and media. Growth media, culture conditions, and genetic manipulations were all as previously described [9]. The yeast strains used in this study are listed in Table 1. A deletion construct to disrupt ATG29 with kanMX was amplified by PCR from BY4741-based atg29 deletion mutants [10]. The resultant cassette was transformed into SEY6210, BJ2169, and KVY55 strains to make TMK4, TMK368, or TMK17, respectively. TMK372 was made by replacing ATG8 gene with the LEU2 gene [11]. TMK105 was
Identification of ATG29/YPL166w
All previously discovered atg mutants, which are defective in autophagy, show a loss of viability under starvation conditions. Thus, sensitivity to starvation would seem to be a good indicator of autophagy defects [15]. To search for novel genes involved in autophagy, we performed a genome-wide screen using a collection of roughly 4500 yeast single-gene deletion mutants in the BY4741 background [10]. Each strain was first screened for the reduced viability under starvation conditions and over
Discussion
The yeast, S. cerevisiae, represents an excellent system for studying the molecular mechanisms of autophagy. In this organism, the machinery used in autophagy is similar to that used by the Cvt pathway. Among the 27 ATG genes involved in the Cvt pathway and/or autophagy, ATG17 is the only gene known to function specifically in autophagy. Recently, novel ATG genes required only for the Cvt pathway were isolated by screening for defects in proAPI maturation [12], [22]. In contrast, it is more
Acknowledgments
We thank the members of the Ohsumi laboratory for helpful discussion, and the National Institute for Basic Biology Center for Analytical Instruments for technical assistance. This work was supported in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
References (27)
- et al.
A unified nomenclature for yeast autophagy-related genes
Dev. Cell
(2003) - et al.
Isolation and characterization of autophagy-defective mutants of Saccharomyces cerevisiae
FEBS Lett.
(1993) - et al.
Novel system for monitoring autophagy in the yeast Saccharomyces cerevisiae
Biochem. Biophys. Res. Commun.
(1995) - et al.
Tor, a phosphatidylinositol kinase homologue, controls autophagy in yeast
J. Biol. Chem.
(1998) - et al.
Mai1p is essential for maturation of proaminopeptidase I but not for autophagy
FEBS Lett.
(2002) - et al.
Ald6p is a preferred target for autophagy in yeast, Saccharomyces cerevisiae
J. Biol. Chem.
(2004) - et al.
Studies of cargo delivery to the vacuole mediated by autophagosomes in Saccharomyces cerevisiae
Dev. Cell
(2002) - et al.
Autophagy in yeast demonstrated with proteinase-deficient mutants and conditions for its induction
J. Cell Biol.
(1992) - et al.
Vacuolar import of proteins and organelles from the cytoplasm
Annu. Rev. Cell Dev. Biol.
(1999) - et al.
Ultrastructural analysis of the autophagic process in yeast: detection of autophagosomes and their characterization
J. Cell Biol.
(1994)
The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation
Embo J.
Aminopeptidase I ofSaccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway
J. Cell Biol.
Two distinct pathways for targeting proteins from the cytoplasm to the vacuole/lysosome
J. Cell Biol.
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Abbreviations: PAS, pre-autophagosomal structure; ATG, autophagy-related genes; Cvt, cytoplasm to vacuole targeting; API, aminopeptidase I; GFP, green fluorescent protein; Vps, vacuolar protein-sorting; PE, phosphatidylethanolamine.