Figure 5.

One-hybrid analysis of FAX-1 and NHR-67 DNA binding activity. A. β-galactosidase activity in one-hybrid experiments for yeast containing DR1A binding sites. All strains contained a reporter plasmid derived from pLacZi that contained a single DR1A binding site upstream of the lacZ gene. Each strain included either no activator or a fusion construct containing a nematode nuclear receptor DBD fused to the yeast GAL4 activation domain. B. β-galactosidase activity for yeast containing DR1G binding sites. Error bars show standard deviations. Asterisks indicate results that are significantly different than no activator control by student's t-test (p < 0.05). The difference between the FAX-1 and FAX-1 N19D mutant activators on DR1A sites is also statistically significant. We performed equivalent experiments using strains containing negative control DRNC sites and activator constructs. These strains did not show β-gal activity relative to controls that had no activator (data not shown). We performed equivalent experiments using strains containing MON1 sites and HRWS sites (both AAGTCA monomers) and a FAX-1 DBD activator, which also did not show β-gal activity relative to controls (data not shown).