Methods for functional analysis of macroautophagy in filamentous fungi.
Autophagy is a bulk degradative process responsible for the turnover of membranes, organelles, and proteins in eukaryotic cells. Genetic and molecular regulation of autophagy has been independently elucidated in budding yeast and mammalian cells. In filamentous fungi, autophagy is required for several important physiological functions, such as asexual and sexual differentiation, pathogenic development, starvation stress and programmed cell death during heteroincompatibility. Here, we detail biochemical and microscopy methods useful for measuring the rate of induction of autophagy in filamentous fungi, and we summarize the methods that have been routinely used for monitoring macroautophagy in both yeast and filamentous fungi. The role of autophagy in carbohydrate catabolism and cell survival is discussed along with the specific functions of macroautophagy in fungal development and pathogenesis.
Deng
Yi Zhen
YZ
Fungal Patho-Biology Group, Temasek Life Sciences Laboratory and Department of Biological Sciences, National University of Singapore, Singapore.
Ramos-Pamplona
Marilou
M
Naqvi
Naweed I
NI
D000818:Animals; D001343:Autophagy / Q000502:physiology*; D001681:Biological Assay / Q000379:methods*; D002103:Cadaverine / Q000031:analogs & derivatives / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D005456:Fluorescent Dyes / Q000378:metabolism; D005658:Fungi / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D006003:Glycogen / Q000378:metabolism; D008853:Microscopy / Q000379:methods; D010588:Phagosomes / Q000378:metabolism / Q000648:ultrastructure; D040901:Proteomics / Q000379:methods
D005456:Fluorescent Dyes; D006003:Glycogen; C008542:monodansylcadaverine; D002103:Cadaverine