PL EN


Preferences help
enabled [disable] Abstract
Number of results
2011 | 58 | 3 | 365-374
Article title

Human hAtg2A protein expressed in yeast is recruited to preautophagosomal structure but does not complement autophagy defects of atg2Δ strain

Content
Title variants
Languages of publication
EN
Abstracts
EN
Yeast ScAtg2, an autophagy-related protein, is highly conserved in other fungi and has two homologues in humans, one of which is hAtg2A encoded by the hATG2A/KIAA0404 gene. Region of homology between Atg2 and hAtg2A proteins comprises the C-terminal domain. We used yeast atg2D strain to express the GFP-KIAA0404 gene, its fragment or fusions with yeast ATG2, and study their effects on autophagy. The GFP-hAtg2A protein localized to punctate structures, some of which colocalized with Ape1-RFP-marked preautophagosomal structure (PAS), but it did not restore autophagy in atg2Δ cells. N-terminal fragment of Atg2 and N-terminal fragment of hAtg2A were sufficient for PAS recruitment but were not sufficient to function in autophagy. Neither a fusion of the N-terminal fragment of hAtg2A with C-terminal domain of Atg2 nor a reciprocal fusion were functional in autophagy. hAtg2A, in contrast to yeast Atg2, did not show interaction with the yeast autophagy protein Atg9 but both Atg2 proteins showed interaction with Atg18, a phospholipid-binding protein, in two-hybrid system. Moreover, deletion of ATG18 abrogated PAS recruitment of hAtg2A. Our results show that human hAtg2A can not function in autophagy in yeast, however, it is recruited to the PAS, possibly due to the interaction with Atg18.
Publisher

Year
Volume
58
Issue
3
Pages
365-374
Physical description
Dates
published
2011
received
2011-03-05
revised
2011-06-02
accepted
2011-08-02
(unknown)
2011-08-29
Contributors
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
author
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
  • Institute of Biochemistry and Biophysics, Polish Academy of Sciences, Warszawa, Poland
References
  • Altschul SF, Madden TL, Schaffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25: 3389-3402.
  • Baggett JJ, D'Aquino KE, Wendland B (2003) The Sla2p talin domain plays a role in endocytosis in Saccharomyces cerevisiae. Genetics 165: 1661-1674.
  • Behrends C, Sowa ME, Gygi SP, Harper JW (2010) Network organization of the human autophagy system. Nature 466: 68-76.
  • Bergamini E, Cavallini G, Donati A, Gori Z (2004) The role of macroautophagy in the ageing process, anti-ageing intervention and age-associated diseases. Int J Biochem Cell Biol 36: 2392-2404.
  • Foury F (1997) Human genetic diseases: a cross-talk between man and yeast. Gene 195: 1-10.
  • Goldberg AL (2003) Protein degradation and protection against misfolded or damaged proteins. Nature 426: 895-899.
  • Gozuacik D, Kimchi A (2004) Autophagy as a cell death and tumor suppressor mechanism. Oncogene 23: 2891-2906.
  • Hall TA (1999) BioEdit: a user firendly biological sequence alignment editor and analysis program from Windows 95/98/NT. Nucleis Acids Symposium Series 41: 95-98.
  • Hershko A, Ciechanover A (1998) The ubiquitin system. Annu Rev Biochem 67: 425-479.
  • James P, Halladay J, Craig EA (1996) Genomic libraries and a host strain designed for highly efficient two-hybrid selection in yeast. Genetics 144: 1425-1436.
  • Kabeya Y, Kawamata T, Suzuki K, Ohsumi Y (2007) Cis1/Atg31 is required for autophagosome formation in Saccharomyces cerevisiae. Biochem Biophys Res Commun 356: 405-410.
  • Katoh K, Misawa K, Kuma K, Miyata T (2002) MAFFT: a novel method for rapid multiple sequence alignment based on fast Fourier transform. Nucleic Acids Res 30: 3059-3066.
  • Kawamata T, Kamada Y, Suzuki K, Kuboshima N, Akimatsu H, Ota S, Ohsumi M, Ohsumi Y (2005) Characterization of a novel autophagy-specific gene, ATG29. Biochem Biophys Res Commun 338: 1884-1889.
  • Klionsky DJ, Cueva R, Yaver DS (1992) Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway. J Cell Biol 119: 287-299.
  • Klionsky DJ, Ohsumi Y (1999) Vacuolar import of proteins and organelles from the cytoplasm. Annu Rev Cell Dev Biol 15: 1-32.
  • Klionsky DJ, Cregg JM, Dunn WA Jr, Emr SD, Sakai Y, Sandoval IV, Sibirny A, Subramani S, Thumm M, Veenhuis M, Ohsumi Y (2003) A unified nomenclature for yeast autophagy-related genes. Dev Cell 5: 539-545.
  • Kusama Y, Sato K, Kimura N, Mitamura J, Ohdaira H, Yoshida K (2009) Comprehensive analysis of expression pattern and promoter regulation of human autophagy-related genes. Apoptosis 14: 1165-1175.
  • Lynch-Day MA, Klionsky DJ (2010) The Cvt pathway as a model for selective autophagy. FEBS Lett 584: 1359-1366.
  • Mari M, Reggiori F (2007) Atg9 trafficking in the yeast Saccharomyces cerevisiae. Autophagy 3: 145-148.
  • Mari M, Griffith J, Rieter E, Krishnappa L, Klionsky DJ, Reggiori F (2010) An Atg9-containing compartment that functions in the early steps of autophagosome biogenesis. J Cell Biol 190: 1005-1022.
  • Mumberg D, Muller R, Funk M (1995) Yeast vectors for the controlled expression of heterologous proteins in different genetic backgrounds. Gene 156: 119-122.
  • Nair U, Cao Y, Xie Z, Klionsky DJ (2010) Roles of the lipid-binding motifs of Atg18 and Atg21 in the cytoplasm to vacuole targeting pathway and autophagy. J Biol Chem 285: 11476-11488.
  • Niedenthal RK, Riles L, Johnston M, Hegemann JH (1996) Green fluorescent protein as a marker for gene expression and subcellular localization in budding yeast. Yeast 12: 773-786.
  • Noda T, Matsuura A, Wada Y, Ohsumi Y (1995) Novel system for monitoring autophagy in the yeast Saccharomyces cerevisiae. Biochem Biophys Res Commun 210: 126-132.
  • Obara K, Noda T, Niimi K, Ohsumi Y (2008a) Transport of phosphatidylinositol 3-phosphate into the vacuole via autophagic membranes in Saccharomyces cerevisiae. Genes Cells 13: 537-547.
  • Obara K, Sekito T, Niimi K, Ohsumi Y (2008b) The Atg18-Atg2 complex is recruited to autophagic membranes via phosphatidylinositol 3-phosphate and exerts an essential function. J Biol Chem 283: 23972-23980.
  • Petranovic D, Nielsen J (2008) Can yeast systems biology contribute to the understanding of human disease? Trends Biotechnol 26: 584-590.
  • Reggiori F, Klionsky DJ (2002) Autophagy in the eukaryotic cell. Eukaryot Cell 1: 11-21.
  • Reggiori F, Tucker KA, Stromhaug PE, Klionsky DJ (2004) The Atg1-Atg13 complex regulates Atg9 and Atg23 retrieval transport from the pre-autophagosomal structure. Dev Cell 6: 79-90.
  • Reggiori F, Shintani T, Nair U, Klionsky DJ (2005) Atg9 cycles between mitochondria and the pre-autophagosomal structure in yeasts. Autophagy 1: 101-109.
  • Sherman F (2002) Getting started with yeast. Methods Enzymol 350: 3-41.
  • Shintani T, Suzuki K, Kamada Y, Noda T, Ohsumi Y (2001) Apg2p functions in autophagosome formation on the perivacuolar structure. J Biol Chem 276: 30452-30460.
  • Suzuki K, Ohsumi Y (2010) Current knowledge of the pre-autophagosomal structure (PAS). FEBS Lett 584: 1280-1286.
  • Suzuki K, Kirisako T, Kamada Y, Mizushima N, Noda T, Ohsumi Y (2001) The pre-autophagosomal structure organized by concerted functions of APG genes is essential for autophagosome formation. Embo J 20: 5971-5981.
  • Suzuki K, Kubota Y, Sekito T, Ohsumi Y (2007) Hierarchy of Atg proteins in pre-autophagosomal structure organization. Genes Cells 12: 209-218.
  • van der Vaart A, Mari M, Reggiori F (2008) A picky eater: exploring the mechanisms of selective autophagy in human pathologies. Traffic 9: 281-289.
  • Voth WP, Jiang YW, Stillman DJ (2003) New 'marker swap' plasmids for converting selectable markers on budding yeast gene disruptions and plasmids. Yeast 20: 985-993.
  • Wang AY, Schulze JM, Skordalakes E, Gin JW, Berger JM, Rine J, Kobor MS (2009) Asf1-like structure of the conserved Yaf9 YEATS domain and role in H2A.Z deposition and acetylation. Proc Natl Acad Sci USA 106: 21573-21578.
  • Wang CW, Kim J, Huang WP, Abeliovich H, Stromhaug PE, Dunn WA Jr, Klionsky DJ (2001) Apg2 is a novel protein required for the cytoplasm to vacuole targeting, autophagy, and pexophagy pathways. J Biol Chem 276: 30442-30451.
  • Xie Z, Klionsky DJ (2007) Autophagosome formation: core machinery and adaptations. Nat Cell Biol 9: 1102-1109.
Document Type
Publication order reference
Identifiers
YADDA identifier
bwmeta1.element.bwnjournal-article-abpv58p365kz
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.