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Shared and specific functions of Arf G proteins at the Golgi

Pennauer, Mirjam Vera. Shared and specific functions of Arf G proteins at the Golgi. 2021, Doctoral Thesis, University of Basel, Faculty of Science.

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Abstract

The ADP-ribosylation factors (Arfs) are small GTPases regulating membrane traffic in the secretory pathway. They are closely related and appear to have overlapping functions, regulators, and effectors. The functional specificity of individual Arfs and the extent of redundancy in vivo are still largely unknown. We addressed these questions by CRISPR/Cas9-mediated genomic deletion of the human class I (Arfs 1 and 3) and class II (Arfs 4 and 5) Arfs, either individually or in combination. Cells lacking individual Arfs or certain combination thereof were viable with only a slight growth defect when lacking Arf1 or Arf4, whereas the combinations of Arf1 and 4, and Arf4 and 5 could not be deleted simultaneously. Hence, class I Arfs are not essential and Arf4 alone was found to be sufficient for cell viability. Remarkably, two single knockouts produced specific and distinct phenotypes. Upon deletion of Arf1, the Golgi complex was enlarged and recruitment of vesicle coats decreased, confirming a major role of Arf1 in coat formation at the Golgi. Cell lines deleted for Arf4 exhibited secretion of ER resident proteins, indicating a specific defect in coatomer-dependent ER protein retrieval by the KDEL receptors. The knockout cell lines will be a useful tool to study other Arf-dependent processes.
Advisors:Spiess, Martin and Spang, Anne and Wieland, Felix
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Growth & Development > Biochemistry (Spiess)
UniBasel Contributors:Spiess, Martin and Spang, Anne
Item Type:Thesis
Thesis Subtype:Doctoral Thesis
Thesis no:14387
Thesis status:Complete
Number of Pages:67
Language:English
Identification Number:
  • urn: urn:nbn:ch:bel-bau-diss143877
edoc DOI:
Last Modified:20 Oct 2021 04:30
Deposited On:19 Oct 2021 14:37

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