Phosphatidylinositol-3-Phosphate Clearance Plays a Key Role in Autophagosome Completion

被引:120
作者
Cebollero, Eduardo [1 ,2 ]
van der Vaart, Aniek [2 ]
Zhao, Mantong [3 ]
Rieter, Ester [2 ]
Klionsky, Daniel J. [3 ]
Helms, J. Bernd [1 ]
Reggiori, Fulvio [2 ]
机构
[1] Univ Utrecht, Dept Biochem & Cell Biol, NL-3508 TD Utrecht, Netherlands
[2] Univ Med Ctr Utrecht, Dept Cell Biol, NL-3584 CX Utrecht, Netherlands
[3] Univ Michigan, Inst Life Sci, Ann Arbor, MI 48109 USA
关键词
SACCHAROMYCES-CEREVISIAE; CVT PATHWAY; YEAST; VACUOLE; DOMAIN; PHOSPHATASES; BIOGENESIS; CYTOPLASM; TRANSPORT; ENDOSOME;
D O I
10.1016/j.cub.2012.06.029
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Background: The biogenesis of autophagosomes, the hallmark of autophagy, depends on the function of the autophagy-related (Atg) proteins and the generation of phosphatidylinositol-3-phosphate (PtdIns3P) at the phagophore assembly site (PAS), the location where autophagosomes arise. The current model is that PtdIns3P is involved primarily in the recruitment of Atg proteins to the PAS and that once an autophagosome is complete, the Atg machinery is released from its surface back into the cytoplasm and reused for the formation of new vesicles. Results: We have identified a PtdIns3P phosphatase, Ymr1, that is essential for the normal progression of both bulk and selective types of autophagy. This protein is recruited to the PAS at an early stage of formation of this structure through a process that requires both its GRAM domain and its catalytic activity. In the absence of Ymr1, Atg proteins fail to dissociate from the limiting membrane of autophagosomes, and these vesicles accumulate in the cytoplasm. Conclusions: Our data thus reveal a key role for PtdIns3P turnover in the regulation of the late steps of autophagosome biogenesis and indicate that the disassembly of the Atg machinery from the surface of autophagosomes is a requisite for their fusion with the vacuole.
引用
收藏
页码:1545 / 1553
页数:9
相关论文
共 38 条
[1]   Autophagy and the cytoplasm to vacuole targeting pathway both require Aut10p [J].
Barth, H ;
Meiling-Wesse, K ;
Epple, UD ;
Thumm, M .
FEBS LETTERS, 2001, 508 (01) :23-28
[2]   BIOCHEMICAL METHODS TO MONITOR AUTOPHAGY-RELATED PROCESSES IN YEAST [J].
Cheong, Heesun ;
Klionsky, Daniel J. .
AUTOPHAGY: LOWER EUKARYOTES AND NON-MAMMALIAN SYSTEMS, PT A, 2008, 451 :1-26
[3]   Specificity of the myotubularin family of phosphatidylinositol-3-phosphatase is determined by the PH/GRAM domain [J].
Choudhury, Papiya ;
Srivastava, Shekhar ;
Li, Zhai ;
Ko, Kyung ;
Albaqumi, Mamdouh ;
Narayan, Kartik ;
Coetzee, William A. ;
Lemmon, Mark A. ;
Skolnik, Edward Y. .
JOURNAL OF BIOLOGICAL CHEMISTRY, 2006, 281 (42) :31762-31769
[4]   A multispecificity syntaxin homologue, Vam3p, essential for autophagic and biosynthetic protein transport to the vacuole [J].
Darsow, T ;
Rieder, SE ;
Emr, SD .
JOURNAL OF CELL BIOLOGY, 1997, 138 (03) :517-529
[5]   Zebrafish MTMR14 is required for excitation-contraction coupling, developmental motor function and the regulation of autophagy [J].
Dowling, J. J. ;
Low, S. E. ;
Busta, A. S. ;
Feldman, E. L. .
HUMAN MOLECULAR GENETICS, 2010, 19 (13) :2668-2681
[6]   Autophagy-based unconventional secretory pathway for extracellular delivery of IL-1β [J].
Dupont, Nicolas ;
Jiang, Shanya ;
Pilli, Manohar ;
Ornatowski, Wojciech ;
Bhattacharya, Dhruva ;
Deretic, Vojo .
EMBO JOURNAL, 2011, 30 (23) :4701-4711
[7]   Quantitative analysis of autophagy-related protein stoichiometry by fluorescence microscopy [J].
Geng, Jiefei ;
Baba, Misuzu ;
Nair, Usha ;
Klionsky, Daniel J. .
JOURNAL OF CELL BIOLOGY, 2008, 182 (01) :129-140
[8]   Cvt18/Gsa12 is required for cytoplasm-to-vacuole transport, pexophagy, and autophagy in Saccharomyces cerevisiae and Pichia pastoris [J].
Guan, J ;
Stromhaug, PE ;
George, MD ;
Habibzadegah-Tari, P ;
Bevan, A ;
Dunn, WA ;
Klionsky, DJ .
MOLECULAR BIOLOGY OF THE CELL, 2001, 12 (12) :3821-3838
[9]   Endosome maturation [J].
Huotari, Jatta ;
Helenius, Ari .
EMBO JOURNAL, 2011, 30 (17) :3481-3500
[10]  
Hutchins MU, 1999, J CELL SCI, V112, P4079