Table 2.

RNA binding of AUF1 p45 and variants to flaviviral RNA annealing substrates.

	5ʹCS			3ʹCS
Protein	KD (nM)a	KD ratiob	ΔΔG (kJ/mol)c	KD (nM)a	KD ratiob	ΔΔG (kJ/mol)c
AUF1 p45 WT	105 ± 27	1	0	101 ± 30	1	0
AUF1 p45 RGG5K	208 ± 19	2	1.7	198 ± 34	2	1.7
AUF1 p45 RGG5A	2157 ± 261	21	7.5	1813 ± 432	18	7.1
AUF1 p45 R272A	230 ± 18	2.2	1.9	201 ± 31	2	1.7
AUF1 p45 R278A	257 ± 51	2.5	2.2	242 ± 67	2.4	2.2
AUF1 p45 R280A	234 ± 56	2.2	1.9	228 ± 47	2.3	2
AUF1 p45 R282A	244 ± 60	2.3	2	234 ± 35	2.3	2
AUF1 p45 R345A	144 ± 5	1.4	0.8	139 ± 17	1.4	0.8

^aDissociation constants and standard deviations derived from at least three measurements.

^bK_D ratio to compare binding of wildtype (WT) vs protein variant.

^cThe binding free energy difference between a WT-RNA complex and a variant-RNA complex was calculated as follows: ΔΔG = -RT ln(K_D WT/K_D variant).