Tonks, Adam; Domgaard, Hannah; Crowley, Valerie; Neumann, Gina; Keiser, Dylan; Metcalf, Josie; Guo, Hongjie; Zhou, Yi; Begemann, Mathew; Taylor, David; Jackson, Ryan (Utah State University)
Faculty Advisor: Jackson, Ryan (College of Science, Chemistry and Biochemistry)
Cms1 is a Type V endonuclease that contains a novel domain, shares little sequence homology with other Type V endonucleases, and in some organisms, is found near genes coding for other single-subunit nucleases. Studies in rice (Oryza sativa) have shown Cms1 capable of RNA-directed DNA editing. However, the mechanism of DNA cleavage remains unknown.
Here we present biochemical data that demonstrate Cms1 from Sulfuricurvum processes an RNA guide and binds/cleaves single- and double-stranded DNA through RuvC nuclease motifs. 2-D classification of structures obtained by negative staining electron microscopy show a major conformational change between SuCms1 bound and unbound to an RNA guide. The predicted global structure appears to be different than those reported for other Type V effectors. These data provide for a greater understanding of Type V endonucleases and may provide an alternative tool for genome editing applications.