Extended Data Fig. 6: Structure determination of AriA purified using a Ocr-Strep pulldown.
From: A virally encoded tRNA neutralizes the PARIS antiviral defence system
Image processing and analysis was performed in cryoSPARC v4.41. a, Representative micrograph from a total of 10,340 movies. Micrographs with CTF-fits worse than 8 Å-resolution were discarded. Particle picking on 9,399 micrographs identified 4,415,782 particles which were extracted and subjected to 2-D Classification. b, Selected 2-D classes corresponding to the AriA hexamer contained 667,479 particles. c, 2-Class ab initio reconstruction of particles from panel b. The gray class corresponds to the AriA homohexamer, while particles associated with the orange class were discarded. Density for the ATPase domains is present, but one blade of the propeller is poorly resolved due to the increased flexibility of the AriA scaffold relative to the fully assembled PARIS complex. d, After iterative rounds of sorting the 392,989 particles from panel C, a stack containing 62,732 particles was isolated and C3 refinement produced a 4.4 Å-resolution reconstruction of the AriA scaffold.
