Function of the SNARE Ykt6 on autophagosomes requires the Dsl1 complex and the Atg1 kinase complex. (7th October 2020)

• Record Type: Journal Article
• Authors: Gao, Jieqiong; Kurre, Rainer; Rose, Jaqueline; Walter, Stefan; Fröhlich, Florian; Piehler, Jacob; Reggiori, Fulvio; Ungermann, Christian
• Is Part Of: EMBO reports. Volume 21:Number 12(2020)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2020-10-07
• Subjects: autophagy -- COPII vesicles -- Dsl1 complex -- SNARE -- Ykt6; Molecular biology -- Periodicals; Molecular Biology -- Periodicals; Molecular biology; Periodicals; 572.8
• DOI: 10.15252/embr.202050733
• Languages: English
• ISSNs: 1469-221X

Abstract

Abstract: The mechanism and regulation of fusion between autophagosomes and lysosomes/vacuoles are still only partially understood in both yeast and mammals. In yeast, this fusion step requires SNARE proteins, the homotypic vacuole fusion and protein sorting (HOPS) tethering complex, the RAB7 GTPase Ypt7, and its guanine nucleotide exchange factor (GEF) Mon1‐Ccz1. We and others recently identified Ykt6 as the autophagosomal SNARE protein. However, it has not been resolved when and how lipid‐anchored Ykt6 is recruited onto autophagosomes. Here, we show that Ykt6 is recruited at an early stage of the formation of these carriers through a mechanism that depends on endoplasmic reticulum (ER)‐resident Dsl1 complex and COPII‐coated vesicles. Importantly, Ykt6 activity on autophagosomes is regulated by the Atg1 kinase complex, which inhibits Ykt6 through direct phosphorylation. Thus, our findings indicate that the Ykt6 pool on autophagosomal membranes is kept inactive by Atg1 phosphorylation, and once an autophagosome is ready to fuse with vacuole, Ykt6 dephosphorylation allows its engagement in the fusion event. Synopsis: The SNARE Ykt6 required the Dsl1 complex to be recruited to autophagosomes. There, it is kept inactive by Atg1 kinase phosphorylation until autophagosomes are ready to fuse with vacuoles. Mutants in Dsl1 complex subunits impair Ykt6 recruitment to autophagosomes. Atg1 kinase directly phosphorylates Ykt6 within its SNARE domain. Atg1 controls the fusogenic activity of Ykt6. Abstract : The SNARE Ykt6 required the Dsl1 complex to be recruited to autophagosomes. There, it is kept inactive by Atg1 kinase phosphorylation until autophagosomes are ready to fuse with vacuoles.