sirolimus and Autolysis

Autophagy and autolysis are two cannibalistic pathways which allow filamentous fungi to obtain nutrients once environmental nutrient sources are exhausted. In Aspergillus nidulans, the effects of mutations in two key autophagy genes, atgA, the ATG1 ortholog, and atgH, the ATG8 ortholog, were compared with mutations in xprG, which encodes a transcriptional activator that plays a key role in autolysis. The anti-fungal drug rapamycin induces autophagy in a range of organisms. Mutations in atgA and atgH did not alter sensitivity to rapamycin, which inhibits growth and asexual spore production (conidiation), indicating that autophagy is not required for rapamycin sensitivity in A. nidulans. In contrast, inhibition of conidiation by rapamcyin was partially suppressed by the xprG1 gain-of-function mutation, indicating that XprG acts in the pathway(s) affected by rapamycin. It was anticipated that the absence of an intact autophagy pathway would accelerate the response to starvation. However, extracellular and intracellular protease production in response to carbon or nitrogen starvation was not increased in the atgAΔ and atgHΔ mutants, and the onset of autolysis was not accelerated. Compared to wild-type strains and the xprGΔ and xprG1 mutants, conidiation of the autophagy mutants was reduced in carbon- or nitrogen-limiting conditions but not during growth on nutrient-sufficient medium. Nuclear localization of the global nitrogen regulator AreA in response to nitrogen starvation was blocked in the xprG2 loss-of-function mutant, but not in the atgHΔ mutant. Conversely, the atgAΔ mutation but not the xprGΔ mutation prevented vacuolar accumulation of GFP-AtgH, a hallmark of autophagy. These results indicate that in A. nidulans there is little interaction between autophagy and autolysis and the two pathways are activated in parallel during starvation.

Topics: Aspergillus nidulans; Autolysis; Autophagy; Carbon; Fungal Proteins; Genes, Fungal; Mutation; Nitrogen; Sirolimus; Starvation; Transcription Factors