Using photoactivatable proteins to monitor autophagosome lifetime
- PMID: 19200874
- DOI: 10.1016/S0076-6879(08)03603-3
Using photoactivatable proteins to monitor autophagosome lifetime
Abstract
Many conditions are now known to cause autophagosome proliferation in cells and organisms including amino acid and serum starvation, ER and oxidative stress, and pathogen infection. Autophagosome proliferation is also observed in disease states and developmental programs. The widespread use of GFP-Atg8 fusion molecules has provided a simple way to visualize the proliferation of autophagosomes in cells. However, GFP-Atg8 markers do not reveal the underlying cause of autophagosome proliferation. Two processes regulate the number of autophagosomes present in cells: (1) formation of the structures and (2) their turnover through fusion with lysosomes. Here we describe the use of photoactivatable proteins to decouple the processes of autophagosome formation from autophagosome turnover. Photoactivatable proteins fused to Atg8 homologs make it possible to pulse-label existing populations of autophagosomes in living cells. The fate of those pulse-labeled autophagosomes can then be monitored to determine autophagosome lifetime. This assay is applicable to both engineered tissue culture models and transgenic organisms expressing photoactivatable proteins fused to Atg8 homologs.
Similar articles
-
In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker.Mol Biol Cell. 2004 Mar;15(3):1101-11. doi: 10.1091/mbc.e03-09-0704. Epub 2003 Dec 29. Mol Biol Cell. 2004. PMID: 14699058 Free PMC article.
-
The amino-terminal region of Atg3 is essential for association with phosphatidylethanolamine in Atg8 lipidation.FEBS Lett. 2009 Apr 2;583(7):1078-83. doi: 10.1016/j.febslet.2009.03.009. Epub 2009 Mar 13. FEBS Lett. 2009. PMID: 19285500
-
A possible role of Atg8 homologs as a scaffold for signal transduction.Autophagy. 2011 Sep;7(9):1080-1. doi: 10.4161/auto.7.9.16178. Epub 2011 Sep 1. Autophagy. 2011. PMID: 21597320
-
Molecular Mechanisms of Autophagy in Plants: Role of ATG8 Proteins in Formation and Functioning of Autophagosomes.Biochemistry (Mosc). 2016 Apr;81(4):348-63. doi: 10.1134/S0006297916040052. Biochemistry (Mosc). 2016. PMID: 27293092 Review.
-
Coordination of Autophagosome-Lysosome Fusion by Atg8 Family Members.Curr Biol. 2018 Apr 23;28(8):R512-R518. doi: 10.1016/j.cub.2018.02.034. Curr Biol. 2018. PMID: 29689234 Review.
Cited by
-
Regulation of autophagy by amino acids and MTOR-dependent signal transduction.Amino Acids. 2015 Oct;47(10):2037-63. doi: 10.1007/s00726-014-1765-4. Epub 2014 Jun 1. Amino Acids. 2015. PMID: 24880909 Free PMC article. Review.
-
Applications of phototransformable fluorescent proteins for tracking the dynamics of cellular components.Photochem Photobiol Sci. 2015 Oct;14(10):1787-806. doi: 10.1039/c5pp00174a. Photochem Photobiol Sci. 2015. PMID: 26345171 Free PMC article. Review.
-
Photoactivatable fluorescent proteins for diffraction-limited and super-resolution imaging.Trends Cell Biol. 2009 Nov;19(11):555-65. doi: 10.1016/j.tcb.2009.09.003. Trends Cell Biol. 2009. PMID: 19836954 Free PMC article. Review.
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Other Literature Sources
Molecular Biology Databases
