In nuclear extract from Cos cells , the affinity of binding of promoter fragment containing a __NODE__ binding site 2 ( 5 ' GGAAAAT 3 ' from __NODE__ gene and __NODE__ protein is greater than the affinity of binding of promoter fragment containing a __NODE__ binding site 2 ( 5 ' GGAAAAT 3 ' from __NODE__ gene and mutant __NODE__ protein ( R474A N475A T541G with its __NODE__ homology domain mutated ) .