Arfaptin-1 negatively regulates Arl1-mediated retrograde transport

PloS One
Lien-Hung HuangChia-Jung Yu

Abstract

The small GTPase Arf-like protein 1 (Arl1) is well known for its role in intracellular vesicular transport at the trans-Golgi network (TGN). In this study, we used differential affinity chromatography combined with mass spectrometry to identify Arf-interacting protein 1b (arfaptin-1b) as an Arl1-interacting protein and characterized a novel function for arfaptin-1 (including the arfaptin-1a and 1b isoforms) in Arl1-mediated retrograde transport. Using a Shiga-toxin subunit B (STxB) transportation assay, we demonstrated that knockdown of arfaptin-1 accelerated the retrograde transport of STxB from the endosome to the Golgi apparatus, whereas Arl1 knockdown inhibited STxB transport compared with control cells. Arfaptin-1 overexpression, but not an Arl1 binding-defective mutant (arfaptin-1b-F317A), consistently inhibited STxB transport. Exogenous arfaptin-1 expression did not interfere with the localization of the Arl1-interacting proteins golgin-97 and golgin-245 to the TGN and vice versa. Moreover, we found that the N-terminal region of arfaptin-1 was involved in the regulation of retrograde transport. Our results show that arfaptin-1 acts as a negative regulator in Arl1-mediated retrograde transport and suggest that different f...Continue Reading

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Citations

Oct 20, 2017·Journal of Virology·Kirstin VondersteinAnna K Överby
May 5, 2017·Journal of Cell Science·Chia-Jung Yu, Fang-Jen S Lee
Apr 29, 2018·Cell Communication and Signaling : CCS·Rae-Mann HsuChia-Jung Yu
Mar 15, 2021·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Hsiang-Pu FengChia-Jung Yu

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Datasets Mentioned

BETA
GM130

Methods Mentioned

BETA
GTPase
differential affinity chromatography
two-hybrid
transfection
pull-down
PCR
gene knockdown

Software Mentioned

Axio vision Rel
MetaMorph
Scaffold proteome

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