Arfaptin‐1 Negatively Regulates Arl1‐mediated Retrograde Transport
| Autor: | Shu-Ting You, Wei-Chung Lee, Lien-Hung Huang, Chia-Jung Yu, Chia-Chen Cheng |
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| Rok vydání: | 2015 |
| Předmět: |
ADP ribosylation factor
Endosome Protein subunit Vesicular Transport Proteins lcsh:Medicine Fluorescent Antibody Technique Biology Autoantigens Shiga Toxin 2 Biochemistry Chromatography Affinity Mass Spectrometry symbols.namesake Genetics Humans Small GTPase lcsh:Science Molecular Biology Adaptor Proteins Signal Transducing Analysis of Variance Gene knockdown Multidisciplinary ADP-Ribosylation Factors Chemistry lcsh:R Golgi Matrix Proteins Membrane Proteins Golgi apparatus Cell biology Vesicular transport protein Gene Knockdown Techniques symbols Axoplasmic transport lcsh:Q RNA Interference Arfaptin-1 Intracellular HeLa Cells trans-Golgi Network Research Article Biotechnology |
| Zdroj: | PLoS ONE PLoS ONE, Vol 10, Iss 3, p e0118743 (2015) |
| ISSN: | 1530-6860 0892-6638 |
| DOI: | 10.1096/fasebj.29.1_supplement.lb179 |
| Popis: | 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 functional complexes containing Arl1 form in distinct microdomains and are responsible for different functions. |
| Databáze: | OpenAIRE |
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