Figure 4.

chANP32A Exhibits Enhanced Interaction with Avian- and Mammalian-Signature vPol Complexes via the SIM-like Sequence and LCAR

(A) 293T cells were transfected with the indicated FLAG-tagged constructs, together with PB1, PA, and PB2 (627E or 627K). Following anti-FLAG precipitation, the indicated proteins were detected by western blot. Representative data from three independent experiments are shown.

(B) Schematic representation of chANP32A. LRR, leucine-rich repeat (repeats 1–5 indicated); LCAR, low-complexity acidic region; SLS, SIM-like sequence; NLS, nuclear localization signal.

(C) Immunofluorescence analysis of the indicated FLAG-chANP32A proteins in transfected MRC5 cells. Scale bars represent 10 μm.

(D) (top) Impact of chANP32A LCAR truncations on PB2-627E vPol activity. Experiment performed as in Figure 1B. (bottom) Western blot analysis of 293T cell lysates from top.

(E and F) Relative impact of chANP32A LCAR truncations (E) and chANP32A-mut3 (F) on PB2-627E vPol interaction. Experiments performed as in (A). Representative data from three independent experiments are shown.

(G) Relative impact of chANP32A LCAR truncations and chANP32A-mut3 on PB2-627E vPol activity. Experiments performed as in Figure 1B.

In (D) and (G), bars represent mean values from three independent experiments (±SD). Significance was determined by Student’s t test (^∗p < 0.05; ^∗∗p < 0.01; ^∗∗∗p < 0.001; ^∗∗∗∗p < 0.0001).