TABLE 1.

Effects of point mutations in P protein on binding to M2-1 and N proteins and minigenome activities

Mutation in P protein	Bindinga to protein:		Minigenome activitya
	M2-1	N	Luciferaseb	CATc
None (WT)	100	100	100	100
Δ100-120	6	184	<2	ND
LY101/102AA	3	126	<2	142
KE103/104AA	263	ND	38	40
TI105/106AA	147	ND	109	ND
E107A	84	ND	ND	ND
TF108/109AA	11	165	25	32
D110A	58	ND	ND	ND
NN111/112AA	111	ND	ND	ND
EE113/114AA	207	ND	171	ND
EE114/115AA	45	ND	110	ND
SS116/117AA	102	ND	104	70
YS118/119AA	57	64	<2	7
S120A	103	124	ND	ND
EE121/122AA	300	103	2	7
L101A	4	227	<2	173
Y102A	4	144	3	223
T108A	77	91	60	64
F109A	3	105	4	97
F241A	125	4	<2	<2
No P	NA	NA	<2	<2
No M2-1	NA	NA	<2	100

^aAll values are relative to WT P, which was set at 100%. ND, not determined; NA, not applicable.

^bEach luciferase minigenome activity value is the average of two to four separate assays, each performed in duplicate (with the exception of P Δ100-120 and with no M2-1 which were each tested only once in duplicate).

^cThe CAT minigenome determinations were each performed twice. The raw data obtained in each of the two assays were similar. The data shown are from only one of the two experiments and the values shown were normalized for relative levels of N and P to account for the decreased levels of WT P (three- to fivefold) observed in Western blots with anti-RSV antibody (BioDesign) and K-109 (polyclonal anti-P). All CAT minigenome determinations were performed in the absence of the M2-1 plasmid.