Atg1 kinase in fission yeast is activated by Atg11-mediated dimerization and cis-autophosphorylation

Pan, Zhao-Qian; Shao, Guang-Can; Liu, Xiao Man; Chen, Quan; Dong, Meng‐Qiu; Du, Li-Lin

doi:10.7554/elife.58073

Cited by 30 publications

(27 citation statements)

References 59 publications

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“…Atg11 and Atg17 were also observed to self-interact in the Pil1 co-tethering assay (Fig. S1E,F, 4B,C,D,E), consistent with previously published results showing that both Atg11 and Atg17 can homodimerize (Nanji et al, 2017;Pan et al, 2020). Together, within the fission yeast Atg1 complex, the Pil1 co-tethering assay recapitulated six previously known binary interactions (Atg1-Atg11, Atg13-Atg17, Atg13-Atg101, Atg1-Atg13, Atg11-Atg11, and Atg17-Atg17), and identified two previously unknown binary interactions (Atg1-Atg17 and Atg11-Atg13).…”

Section: Pil1 Co-tethering Assaysupporting

confidence: 92%

“…S1C,D, S2A,B, 4D,E). These four pairs of interactions are consistent with previously published results obtained using in-vitro pull-down of recombinant proteins (Nanji et al, 2017;Pan et al, 2020), indicating that these pairs of relatively strong interactions identified by the Pil1 co-tethering assay are direct physical interactions.…”

Section: Pil1 Co-tethering Assaysupporting

confidence: 91%

“…The fission yeast Atg1 complex plays important roles in the initiation of starvationinduced autophagy and includes five components, namely, Atg1, Atg11, Atg13, Atg17, and Atg101 (Nanji et al, 2017;Pan et al, 2020;Sun et al, 2013;Suzuki et al, 2015) (Fig. 4A).…”

Section: Pil1 Co-tethering Assaymentioning

confidence: 99%

“…The fission yeast Schizosaccharomyces pombe is a widely-used and powerful model organism for dissecting the mechanisms of a diverse range of cellular processes (Hoffman et al, 2015). For example, in recent years, we and others have used S. pombe to study autophagy (Fukuda et al, 2020;Liu et al, 2018;Matsuhara and Yamamoto, 2016;Mukaiyama et al, 2009;Nanji et al, 2017;Pan et al, 2020;Sun et al, 2013;Suzuki et al, its C-terminal cytosolic tail (Fig. 2D).…”

Section: Introductionmentioning

confidence: 99%

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Visual detection of binary, ternary, and quaternary protein-protein interactions in fission yeast by Pil1 co-tethering assay

Liu

Zhao

et al. 2021

Preprint

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Protein-protein interactions are vital for executing nearly all cellular processes. To facilitate the detection of protein-protein interactions in living cells of the fission yeast Schizosaccharomyces pombe, here we present an efficient and convenient method termed the Pil1 co-tethering assay. In its basic form, we tether a bait protein to mCherry-tagged Pil1, which forms cortical filamentary structures, and examine whether a GFP-tagged prey protein colocalizes with the bait. We demonstrate that this assay is capable of detecting pairwise protein-protein interactions of cytosolic proteins, transmembrane proteins, and nuclear proteins. Furthermore, we show that this assay can be used for detecting not only binary protein-protein interactions, but also ternary and quaternary protein-protein interactions. Using this assay, we systematically characterized the protein-protein interactions in the Atg1 complex and in the phosphatidylinositol 3-kinase (PtdIns3K) complexes and found that Atg38 is incorporated into the PtdIns3K complex I via an Atg38-Vps34 interaction. Our data show that this assay is a useful and versatile tool and should be added to the routine toolbox of fission yeast researchers.

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Section: Pil1 Co-tethering Assaysupporting

confidence: 92%

Section: Pil1 Co-tethering Assaysupporting

confidence: 91%

Section: Pil1 Co-tethering Assaymentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Visual detection of binary, ternary, and quaternary protein-protein interactions in fission yeast by Pil1 co-tethering assay

Liu

Zhao

et al. 2021

Preprint

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“…Previously we showed that ZZ2 and ZZ3 together are sufficient for binding Lap2 and Ape2 (Liu et al, 2015). To systematically map the cargo-interacting regions of Nbr1, we employed an imaging-based method, termed Pil1 co-tethering assay, to examine pairwise protein-protein interactions (Pan et al, 2020). In this assay, we fuse a bait protein to Pil1, which forms filamentary structures.…”

Section: Resultsmentioning

confidence: 99%

Molecular and structural mechanisms of ZZ domain-mediated cargo recognition by autophagy receptor Nbr1

Wang

Zhang

Liu

et al. 2021

Preprint

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In selective autophagy, cargo selectivity is determined by autophagy receptors. However, it remains scarcely understood how autophagy receptors recognize specific protein cargos. In the fission yeast Schizosaccharomyces pombe, a selective autophagy pathway termed Nbr1-mediated vacuolar targeting (NVT) employs Nbr1, an autophagy receptor conserved across eukaryotes including humans, to target cytosolic hydrolases into the vacuole. Here, we identify two new NVT cargos, the mannosidase Ams1 and the aminopeptidase Ape4, that bind competitively to the first ZZ domain of Nbr1 (Nbr1-ZZ1). High-resolution cryo-EM analyses reveal how a single ZZ domain recognizes two distinct protein cargos. Nbr1-ZZ1 not only recognizes the N-termini of cargos via a conserved acidic pocket, similar to other characterized ZZ domains, but also engages additional parts of cargos in a cargo-specific manner. Our findings unveil a single-domain bispecific mechanism of autophagy cargo recognition, elucidate its underlying structural basis, and expand the understanding of ZZ domain-mediated protein-protein interactions.

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Cellular response during cellular starvation: A battle for cellular survivability

Bhowmick,

Biswas,

Mukherjee

2024

Cell Biochemistry & Function

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Cellular starvation occurs when a cell is deprived of nutrition and oxygen availability. The genesis of this state of deprivation is exclusively contingent upon the inadequacy in the supply of essential components, namely amino acids, glucose, and oxygen. Consequently, the impact of this altered condition manifests in the regulation of cellular respiratory, metabolic, and stress responses. Subsequently, as a reactive outcome, cell death may transpire through mechanisms such as autophagy or apoptosis, particularly under prolonged circumstances. However, the cell combats such situations by evolving altered activity in their metabolic and protein level. Modulated signaling cascades help them to conquer starvation. But as in a prolonged condition, the battle that a cell has to evolve will come into and result in the form of cellular death. Therefore, in cancer therapy, cellular starvation may also act as a possible way out so that the cancer cell can undergo its death pathway in an induced starved condition. This review has collectively depicted the mechanism of cellular starvation. Besides this, the cellular response in this starved condition has also been summarized. Gaining such knowledge of the causation of cell starvation and cellular response during starvation not only generates new insight into the mechanism of cell survivability but also may act as a beneficial role in combating cellular diseases like cancer.

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Atg1 kinase in fission yeast is activated by Atg11-mediated dimerization and cis-autophosphorylation

Cited by 30 publications

References 59 publications

Visual detection of binary, ternary, and quaternary protein-protein interactions in fission yeast by Pil1 co-tethering assay

Visual detection of binary, ternary, and quaternary protein-protein interactions in fission yeast by Pil1 co-tethering assay

Molecular and structural mechanisms of ZZ domain-mediated cargo recognition by autophagy receptor Nbr1

Cellular response during cellular starvation: A battle for cellular survivability

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