The Glo3 GAP crystal structure supports the molecular niche model for ArfGAPs in COPI coats

Xie, Boyang; Jung, Christian; Chandra, Mintu; Engel, Andrew G.; Kendall, Amy; Jackson, Lauren P.

doi:10.1016/j.jbior.2020.100781

Cited by 7 publications

(5 citation statements)

References 45 publications

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“…, 2009 ; Xie et al. , 2021 , 2023 ), and that COPI vesicles form for both intra-Golgi and Golgi-to-ER transport earlier in Golgi maturation ( Papanikou et al. , 2015 ; Tojima et al.…”

Section: Resultsmentioning

confidence: 99%

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix

Manzer,

Fromme

2023

MBoC

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Arf GTPases are central regulators of the Golgi complex, which serves as the nexus of membrane trafficking pathways in eukaryotic cells. Arf proteins recruit dozens of effectors to modify membranes, sort cargos, and create and tether transport vesicles, and are therefore essential for orchestrating Golgi trafficking. The regulation of Arf activity is controlled by the action of Arf-GEFs, which activate via nucleotide exchange, and Arf-GAPs, which inactivate via nucleotide hydrolysis. The localization dynamics of Arf GTPases and their Arf-GAPs during Golgi maturation have not been reported. Here we use the budding yeast model to examine the temporal localization of the Golgi Arf-GAPs. We also determine the mechanisms used by the Arf-GAP Age2 to localize to the Golgi. We find that the catalytic activity of Age2 and a conserved sequence in the unstructured C-terminal domain of Age2 are both required for Golgi localization. This sequence is predicted to form an amphipathic helix and mediates direct binding of Age2 to membranes in vitro. We also report the development of a probe for sensing active Arf1 in living cells and use this probe to characterize the temporal dynamics of Arf1 during Golgi maturation.

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“…, 2009 ; Xie et al. , 2021 , 2023 ), and that COPI vesicles form for both intra-Golgi and Golgi-to-ER transport earlier in Golgi maturation ( Papanikou et al. , 2015 ; Tojima et al.…”

Section: Resultsmentioning

confidence: 99%

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix

Manzer,

Fromme

2023

MBoC

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“…1 ). We combine data on this new β′-COP/Glo3 BoCCS interaction with a published Glo3 GAP/Arf1 structural model ( Xie et al, 2021 ) to propose how β′-COP may bind Glo3 and Arf1 on membranes in the presence of cargo ( Fig. 5 ).…”

Section: Discussionmentioning

confidence: 99%

An interaction between β′-COP and the ArfGAP, Glo3, maintains post-Golgi cargo recycling

Xie

Guillem

Date

et al. 2023

Journal of Cell Biology

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The essential COPI coat mediates retrieval of transmembrane proteins at the Golgi and endosomes following recruitment by the small GTPase, Arf1. ArfGAP proteins regulate COPI coats, but molecular details for COPI recognition by ArfGAPs remain elusive. Biochemical and biophysical data reveal how β′-COP propeller domains directly engage the yeast ArfGAP, Glo3, with a low micromolar binding affinity. Calorimetry data demonstrate that both β′-COP propeller domains are required to bind Glo3. An acidic patch on β′-COP (D437/D450) interacts with Glo3 lysine residues located within the BoCCS (binding of coatomer, cargo, and SNAREs) region. Targeted point mutations in either Glo3 BoCCS or β′-COP abrogate the interaction in vitro, and loss of the β′-COP/Glo3 interaction drives Ste2 missorting to the vacuole and aberrant Golgi morphology in budding yeast. These data suggest that cells require the β′-COP/Glo3 interaction for cargo recycling via endosomes and the TGN, where β′-COP serves as a molecular platform to coordinate binding to multiple proteins, including Glo3, Arf1, and the COPI F-subcomplex.

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“…In this work, we identify two new direct binding partners for β’-COP: the Glo3 BoCCS region and Arf1 (Figure 1). We combine data on this new β’-COP/Glo3 BoCCS interaction with a published Glo3 GAP/Arf1 structural model 58 to propose how β’-COP may bind Glo3 and Arf1 on membranes in the presence of cargo (Figure 5). Multiple lines of evidence support this model.…”

Section: Discussionmentioning

confidence: 99%

An interaction between β’-COP and the ArfGAP, Glo3, maintains post-Golgi cargo recycling

Xie

Guillem

Jung

et al. 2022

Preprint

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The essential COPI vesicular coat mediates retrieval of key transmembrane proteins at the Golgi and endosomes following recruitment by the small GTPase, Arf1. ArfGAP proteins regulate COPI coats, but molecular details for COPI recognition by ArfGAP proteins remain elusive. Biochemical and biophysical data reveal how β’-COP propeller domains directly engage the yeast ArfGAP, Glo3, with a low micromolar binding affinity (KD ~1 µM). Calorimetry data demonstrate both β’-COP propeller domains are required to bind Glo3 using electrostatic interactions. An acidic patch on β’-COP (D437/D450) interacts with Glo3 lysine residues located within the BoCCS (Binding of Coatomer, Cargo, and SNAREs) region. Targeted point mutations in either Glo3 BoCCS or β’-COP abrogate the interaction in vitro, and loss of the β’-COP/Glo3 interaction drives Ste2 mis-sorting to the vacuole and aberrant Golgi morphology in budding yeast. Together, these data suggest cells require the β’-COP/Glo3 interaction for cargo recycling via endosomes and the TGN, where β’-COP may serve as a molecular platform to coordinate binding to multiple protein partners, including Glo3, Arf1, and the COPI F-subcomplex.

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The Glo3 GAP crystal structure supports the molecular niche model for ArfGAPs in COPI coats

Cited by 7 publications

References 45 publications

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix

The Arf-GAP Age2 localizes to the late-Golgi via a conserved amphipathic helix

An interaction between β′-COP and the ArfGAP, Glo3, maintains post-Golgi cargo recycling

An interaction between β’-COP and the ArfGAP, Glo3, maintains post-Golgi cargo recycling

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