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PreprintCited by 7Cellular transport and secretionFungal and yeast genetics researchProtein Kinase Regulation and GTPase Signaling

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

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Manzer, Kaitlyn M., Fromme, J. Christopher·2023·Molecular Biology of the Cell
DOI 10.1101/2023.07.23.550229PMC PMC10846627

Preprint — published as 10.1091/mbc.e23-07-0283 · Paper page

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Abstract

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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Metadata

Authors
Manzer, K. M., & Fromme, J. C.
Affiliations
Cornell University (US)
Corresponding author
Fromme, J. Christopher
ORCID
Venue
Molecular Biology of the Cell
Publisher
openRxiv
Published
2023-07-24
Volume
34
Issue
12
Pages
ar119-ar119
ISSN
1059-1524 (ISSN-L)
DOI
10.1101/2023.07.23.550229
PMC
PMC10846627
OpenAlex
W4386475638
Cited by
7
References
1
Citations by year
2026: 3 · 2025: 1 · 2024: 3
FWCI
1.01
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Molecular Biology of the Cell (publishedVersion)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by-nc-sa)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
Funders
  • National Science Foundation
  • National Institutes of Health
  • National Institute of General Medical Sciences (R35GM136258)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
ArfGAP, Arf1, Golgi apparatus, amphipathic helix, Golgi maturation, membrane trafficking, Arf GTPases, ARF-GEF, nucleotide hydrolysis, membrane binding, Saccharomyces cerevisiae

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