Mutational analysis of the rotavirus NSP4 enterotoxic domain that binds to caveolin-1

Table 2 Phenotypes of pD32-NSP4 mutants plus pD22-caveolin-1 ^a

	CSM Leu^-Trp^-His^- + 3AT	CSM Leu^-Trp^-Ura^-	CSM Leu^-Trp^- + 0.2% 5FOA	beta-gal	Phenotypes^c
	50 mM	CSM Leu^-Trp^-Ura^-	CSM Leu^-Trp^- + 0.2% 5FOA	Units^b	Phenotypes^c
Negative Control	+/−	-	-	0.134	Negative
NSP4 46-175	+/−	-	+/−	1.390	Positive
NSP4 Ala	-	-	+/−	1.822	Positive
NSP4 HydroMut	-	+/−	+	0.105	Negative
Rev1	-	-	+/−	3.941	Positive
Rev2	+/−	-	+/−	3.042	Positive
Rev3	+/−	+/−	-	3.195	Positive
Rev1,2 127A	+/−	-	+/−	2.581	Positive
Rev1 124,127A	-	-	+/−	4.044	Positive
Rev2 116A	-	-	+/−	0.165	Negative
Rev2 116,124,127A	+/−	+/−	+/−	0.056	Negative

^aColonies were grown on CSM Leu^-Trp^- and replica plated onto CSM Leu^-Trp^-His with 50 mM 3AT; CSM Leu^-Trp^-Ura^-; and CSM Leu^-Trp^- with 0.2% 5FOA.
^bBeta-galactosidase activity was quantitatively measured by CPRG assays.
^cThe phenotype was determined by combining the results of growth on differential media and beta-galactosidase activity.
The negative control and mutants NSP4 clones that did not interact with caveolin-1 are in bold letters.

ISSN: 1743-422X