Analysis of autophagosome membrane cycling by fluorescence microscopy

Methods in Molecular Biology
Julie E Legakis, Daniel J Klionsky

Abstract

Autophagy is a physiological process functionally linked to cellular dynamics during starvation, cardiomyopathies, neurodegeneration, cellular immunity, and certain cancers. Although nearly 30 autophagy-related (ATG) genes have been identified and characterized, the molecular mechanisms of this process are only partially understood. One aspect of the pathway that has been intensely studied is the identity of the membrane source for newly formed autophagosomes. Although it occurs at a basal level, autophagy is an inducible process. The process of autophagosome formation involves recruitment and delivery of membrane and recycling of Atg proteins. Despite continuing attempts to identify the source of the autophagosome membrane, we are only recently beginning to understand the nature of autophagosome formation and the role of membrane protein cycling in this process. There now exists an assay utilizing fluorescence microscopy to monitor the localization, and therefore the movement, of membrane-associated Atg proteins. We describe here a method that allows visualization of Atg membrane proteins in order to observe their potential source membranes and also to determine the temporal order of action of other Atg proteins with regard to...Continue Reading

Citations

Oct 27, 2009·Biochimica Et Biophysica Acta·Marco Falasca, Tania Maffucci
Jan 6, 2015·Traffic·Verónica A SegarraSandra K Lemmon

❮ Previous
Next ❯

Related Concepts

Related Feeds

Cardiomyopathy

Cardiomyopathy is a disease of the heart muscle, that can lead to muscular or electrical dysfunction of the heart. It is often an irreversible disease that is associated with a poor prognosis. There are different causes and classifications of cardiomyopathies. Here are the latest discoveries pertaining to this disease.

Parkinson's Disease & Autophagy (MDS)

Autophagy leads to degradation of damaged proteins and organelles by the lysosome. Impaired autophagy has been implicated in several diseases. Here is the role of autophagy in Parkinson’s disease.

Autophagy Networks

Autophagy is a lysosomal pathway that involves degradation of proteins and functions in normal growth and pathological conditions, through a series of complex networks. The catabolic process involves delivery of proteins and organelles to the lysosome. Here is the latest research on autophagy networks.

Autophagy & Metabolism

Autophagy preserves the health of cells and tissues by replacing outdated and damaged cellular components with fresh ones. In starvation, it provides an internal source of nutrients for energy generation and, thus, survival. A powerful promoter of metabolic homeostasis at both the cellular and whole-animal level, autophagy prevents degenerative diseases. It does have a downside, however--cancer cells exploit it to survive in nutrient-poor tumors.

Autophagy & Disease

Autophagy is an important cellular process for normal physiology and both elevated and decreased levels of autophagy are associated with disease. Here is the latest research.

Autophagy & Model Organisms

Autophagy is a cellular process that allows degradation by the lysosome of cytoplasmic components such as proteins or organelles. Here is the latest research on autophagy & model organisms