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  • Identification of autophagy as a longevity-assurance mechanism in the aging model Podospora anserina.

Identification of autophagy as a longevity-assurance mechanism in the aging model Podospora anserina.

Autophagy (2014-03-04)
Laura Knuppertz, Andrea Hamann, Francesco Pampaloni, Ernst Stelzer, Heinz D Osiewacz
ABSTRACT

The filamentous ascomycete Podospora anserina is a well-established aging model in which a variety of different pathways, including those involved in the control of respiration, ROS generation and scavenging, DNA maintenance, proteostasis, mitochondrial dynamics, and programmed cell death have previously been demonstrated to affect aging and life span. Here we address a potential role of autophagy. We provide data demonstrating high basal autophagy levels even in strains cultivated under noninduced conditions. By monitoring an N-terminal fusion of EGFP to the fungal LC3 homolog PaATG8 over the lifetime of the fungus on medium with and without nitrogen supplementation, respectively, we identified a significant increase of GFP puncta in older and in nitrogen-starved cultures suggesting an induction of autophagy during aging. This conclusion is supported by the demonstration of an age-related and autophagy-dependent degradation of a PaSOD1-GFP reporter protein. The deletion of Paatg1, which leads to the lack of the PaATG1 serine/threonine kinase active in early stages of autophagy induction, impairs ascospore germination and development and shortens life span. Under nitrogen-depleted conditions, life span of the wild type is increased almost 4-fold. In contrast, this effect is annihilated in the Paatg1 deletion strain, suggesting that the ability to induce autophagy is beneficial for this fungus. Collectively, our data identify autophagy as a longevity-assurance mechanism in P. anserina and as another surveillance pathway in the complex network of pathways affecting aging and development. These findings provide perspectives for the elucidation of the mechanisms involved in the regulation of individual pathways and their interactions.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Methyl viologen dichloride hydrate, 98%
Sigma-Aldrich
Anti-Green Fluorescent Protein (GFP), N-terminal antibody, Mouse monoclonal, clone GSN24, purified from hybridoma cell culture
Roche
DIG DNA Labeling Kit, sufficient for 40 labeling reactions, kit of 1 (7 components), suitable for hybridization