Structural basis for Dicer-like function of an engineered RNase III variant and insights into the reaction trajectory of two-Mg²⁺-ion catalysis.

ABSTRACT: The RNase III family of dsRNA-specific endonucleases is exemplified by prokaryotic RNase III and eukaryotic Rnt1p, Drosha, and Dicer. Structures of Aquifex aeolicus RNase III (AaRNase III) and Saccharomyces cerevisiae Rnt1p (ScRnt1p) show that both enzymes recognize substrates in a sequence-specific manner and propel RNA hydrolysis by two-Mg²⁺-ion catalysis. Previously, we created an Escherichia coli RNase III variant (EcEEQ) by eliminating the sequence specificity via protein engineering and called it bacterial Dicer for the fact that it produces heterogeneous small interfering RNA cocktails. Here, we present a 1.8-Å crystal structure of a postcleavage complex of EcEEQ, representing a reaction state immediately after the cleavage of scissile bond. The struc

SUBMITTER: Dharavath S 

PROVIDER: S-EPMC9291653 | biostudies-literature | 2022 Jan

REPOSITORIES: biostudies-literature