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. 2017 Dec 11;7:17309. doi: 10.1038/s41598-017-17621-6

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Aichi virus 3A protein recruits the PI4KB kinase via ACBD3. (A) 3A protein does not recruit PI4KB directly to GUVs. 250 nM CFP-3A and PI4KB labeled by Alexa488 were added to GUVs containing 5% of DGS-NTA(Ni) and ATTO647N-DOPE (0.1 mol %). The CFP-3A signal is in blue, the PI4KB-Alexa488 signal is in green and the ATTO647 signal is in red. Representative image of three independent experiments. Scale bar = 20 µm. (B) PI4KB is recruited to membranes when ACBD3 is present. CFP-3A, Alexa488 labeled PI4KB and unlabeled ACBD3 (250 nM each) were added to GUVs containing 5% DGS-NTA(Ni) and ATTO647N-DOPE (0.1 mol %). The CFP-3A signal is in blue, the PI4KB-Alexa488 signal in green and the ATTO647 signal is in red. Representative image of three independent experiments. Scale bar = 20 µm. (C) FRAP analysis of the 3A:ACBD3:PI4KB protein complex. A small cross-section of a GUV membrane was intensively bleached by a 405 nm laser and fluorescence recovery after photobleaching (FRAP) of CFP-3A was measured. (D) Schematic representation of the 3A:ACBD3:PI4KB protein complex.