Structural domains in phage Mu transposase: identification of the site-specific DNA-binding domain.

Limited proteolysis of phage Mu transposase with three proteases of differing specificities produced a common pattern of fragmentation. The fragments were mapped by using a combination of immunoblotting and amino acid sequence analysis. Our results suggest that the transposase molecule is organized principally into three domains: an amino-terminal domain of molecular mass 30 kDa, a core region of approximately 35 kDa, and a carboxyl-terminal domain of approximately 10 kDa. The amino-terminal domain has at least two additional sites that are partially accessible to proteases. Filter binding and nuclease protection studies were done to determine the functions of the isolated domains. Site-specific binding to Mu DNA was localized to the amino-terminal domain. The core domain showed nonspecific DNA-binding activity.