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Mapping Critical Residues in ATG11's Coiled-Coil 2 Domain that Block Multiple Interactions and Disrupt Selective Autophagy

Front Cell Dev Biol. 2022-01; 
Mitchell D Meyer, Jasmine Winzeler, Sophia M Taylor, Alex Kilgore, Kimberly Edicha, Chase Chitwood, Zachary Spearin, S K Nadia Rahman Silvia, Ronith Chakraborty, Jesse E Smith, Bridget Kennedy, Carson Zois, Hayley Cawthon, Mukiri Gilruth, Steven K Backues
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Mutagenesis Services … pGAD-ATG11 I569E was custom generated by Genscript. All other pGAD-ATG11 single mutants were generated by site-directed mutagenesis using inverse PCR and InFusion cloning (… Get A Quote

摘要

Selective autophagy is a conserved subcellular process that maintains the health of eukaryotic cells by targeting damaged or toxic cytoplasmic components to the vacuole/lysosome for degradation. A key player in the initiation of selective autophagy in (baker's yeast) is a large adapter protein called Atg11. Atg11 has multiple predicted coiled-coil domains and intrinsically disordered regions, is known to dimerize, and binds and organizes other essential components of the autophagosome formation machinery, including Atg1 and Atg9. We performed systematic directed mutagenesis on the coiled-coil 2 domain of Atg11 in order to map which residues were required for its structure and function. Using yeast-2-hybrid and... More

关键词

Atg1, Atg11, Atg19, Atg9, coimmunoprecipitation, directed mutagenesis, selective autophagy, yeast-2-hybrid