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Shared and specific functions of Arfs 1-5 at the golgi revealed by systematic knockouts

ABSTRACT: DP-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 are still largely unknown. We addressed these questions by CRISPR/Cas9-mediated genomic deletion of the human class I (Arf1/3) and class II (Arf4/5) Arfs, either individually or in combination. Most knockout cell lines were viable with slight growth defects only when lacking Arf1 or Arf4. However, Arf1+4, and Arf4+5 could not be deleted simultaneously. Class I Arfs are non-essential and Arf4 alone is sufficient for viability. Upon Arf1 deletion, the Golgi was enlarged and recruitment of vesicle coats decreased, confirming a major role of Arf1 in vesicle formation at the Golgi. Knockout of Arf4 caused secretion of ER-resident proteins, indicating specific defects in coatomer-dependent ER protein retrieval by KDEL receptors. The knockout cell lines will be useful tools to study other Arf-dependent processes.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Katarzyna Buczak

LAB HEAD: Alexander Schmidt

PROVIDER: PXD028846 | Pride | 2021-11-02

REPOSITORIES: Pride

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			Action	DRS
	001_SecrIII_F20.raw	Raw
	001_SecrIII_F20__F066872_.mzid.gz	Mzid
	001_SecrIII_F20__F066872_.mzid_001_SecrIII_F20__F066872_.MGF	Mzid
	002_SecrIII_F20.raw	Raw
	002_SecrIII_F20__F066873_.mzid.gz	Mzid

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The FAM114A proteins are adaptors for the recycling of Golgi enzymes - Fig4b-E

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The FAM114A proteins are adaptors for the recycling of Golgi enzymes - Fig1E-F

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The FAM114A proteins are adaptors for the recycling of Golgi enzymes - Figu 2SA

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