Figure 2 | Scientific Reports

Figure 2

From: Negative charge and membrane-tethered viral 3B cooperate to recruit viral RNA dependent RNA polymerase 3Dpol

Figure 2

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.

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