Binding with heat shock cognate protein HSC70 fine-tunes the Golgi association of the small GTPase ARL5B

Jaimon, Ebsy; Tripathi, Aashutosh; Khurana, Arohi; Ghosh, Dipanjana; Sugatha, Jini; Datta, Soma

doi:10.1016/j.jbc.2021.101422

Cited by 4 publications

(2 citation statements)

References 65 publications

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“…GEF and GAP proteins were not detected in the current study; however, these may be difficult to label and or pull down. A recent study has identified the heat shock protein, HSC70, as an interactive partner of Arl5b [37]. HSC70 was not detected in our study; however, as these authors showed that HSC70 bound preferentially to the GDP-bound form of Arl5b [37], the absence of HSC70 from the interactive partners of GTP-bound Arl5b(Q70L) in our study is probably not surprising.…”

Section: Genecontrasting

confidence: 50%

Interacting partners of Golgi‐localized small G protein Arl5b identified by a combination of in vivo proximity labelling and GFP‐Trap pull down

et al. 2022

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The small G protein Arl5b is localised on the trans-Golgi network (TGN) and regulates endosomes-to-TGN transport. Here, we combined in vivo and in vitro techniques to map the interactive partners and near neighbours of Arl5b at the TGN, using constitutively active, membrane-bound Arl5b (Q70L)-GFP in stably expressing HeLa cells, and the proximity labelling techniques BioID and APEX2 in parallel with GFP-Trap pull down. From MS analysis, 22 Golgi proteins were identified; 50% were TGN-localised Rabs, Arfs and Arls. The scaffold/tethering factors ACBD3 (GCP60) and PIST (GOPC) were also identified, and we show that Arl5b is required for TGN recruitment of ACBD3. Overall, the combination of in vivo labelling and direct pull downs indicates a highly organised complex of small G proteins on TGN membranes.

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Section: Genecontrasting

confidence: 50%

Interacting partners of Golgi‐localized small G protein Arl5b identified by a combination of in vivo proximity labelling and GFP‐Trap pull down

et al. 2022

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“…If necessary, myristoylated Arf6 can be further enriched by ammonium sulfate precipitation. In vitro myristoylation has also been applied to obtain modified Arl3 [68] and Arl5B [69], as well as the myristoylated SRC family kinases [70].…”

Section: Why and How To Prepare Lipid-modified Proteinsmentioning

confidence: 99%

In vitro reconstitution of small GTPase regulation

et al. 2022

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Small GTPases play essential roles in the organization of eukaryotic cells. In recent years, it has become clear that their intracellular functions result from intricate biochemical networks of the GTPase and their regulators that dynamically bind to a membrane surface. Due to the inherent complexities of their interactions, however, revealing the underlying mechanisms of action is often difficult to achieve from in vivo studies. This review summarizes in vitro reconstitution approaches developed to obtain a better mechanistic understanding of how small GTPase activities are regulated in space and time.

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ARMH3 is an ARL5 effector that promotes PI4KB-catalyzed PI4P synthesis at the trans-Golgi network

Ishida,

Golding,

Keren-Kaplan

et al. 2024

Nat Commun

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ARL5 is a member of the ARF family of small GTPases that is recruited to the trans-Golgi network (TGN) by another ARF-family member, ARFRP1, in complex with the transmembrane protein SYS1. ARL5 recruits its effector, the multisubunit tethering complex GARP, to promote SNARE-dependent fusion of endosome-derived retrograde transport carriers with the TGN. To further investigate the function of ARL5, we sought to identify additional effectors. Using proximity biotinylation and protein interaction assays, we found that the armadillo-repeat protein ARMH3 (C10orf76) binds to active, but not inactive, ARL5, and that it is recruited to the TGN in a SYS1-ARFRP1-ARL5-dependent manner. Unlike GARP, ARMH3 is not required for the retrograde transport of various cargo proteins. Instead, ARMH3 functions to activate phosphatidylinositol 4-kinase IIIβ (PI4KB), accounting for the main pool of phosphatidylinositol 4-phosphate (PI4P) at the TGN. This function contributes to recruitment of the oncoprotein GOLPH3 and glycan modifications at the TGN. These studies thus identify the SYS1-ARFRP1-ARL5-ARMH3 axis as a regulator of PI4KB-dependent generation of PI4P at the TGN.

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Binding with heat shock cognate protein HSC70 fine-tunes the Golgi association of the small GTPase ARL5B

Cited by 4 publications

References 65 publications

Interacting partners of Golgi‐localized small G protein Arl5b identified by a combination of in vivo proximity labelling and GFP‐Trap pull down

Interacting partners of Golgi‐localized small G protein Arl5b identified by a combination of in vivo proximity labelling and GFP‐Trap pull down

In vitro reconstitution of small GTPase regulation

ARMH3 is an ARL5 effector that promotes PI4KB-catalyzed PI4P synthesis at the trans-Golgi network

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