The protein splicing domain of the homing endonuclease PI-SceI is responsible for specific DNA binding

摘要

The homing endonuclease PI-SceI consists of a protein splicing domain (I) and an endonucleolytic domain (II). To characterize the two domains with respect to their contribution to DNA recognition we cloned, purified and characterized the isolated domains. Both domains have no detectable endonucleolytic activity. Domain I binds specifically to the PI-SceI recognition sequence, whereas domain II displays only weak non-specific DNA binding. In the specific complex with domain I the DNA is bent to a similar extent as observed with the initial complex formed between PI-SceI and DNA. Our results indicate that protein splicing domain I is also involved in recognition of the DNA substrate.