Researches on ribozymes at molecular and atomic level

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The molecular design, structure and reaction mechanism of the naturally occurring ribozymes were investigated. The minimal catalytic unit of Group I intron ribozymes was identified. The RNA folding pathways and the intermolecular long-range interactions of the intron were identified. In vitro selection te4chnique was successfully employed for producing artificial ribozymes and RNPs. Hammerhead ribozyme was engineered successfully for in vivo use. New gene discovery system was developed. Ribosomal RNAs of mammalian mitochondria and the corresponding protein components were identified and analyzed. It was elucidated that the binding of stalk proteins to rRNA in the ribosomal GTPase center is involved in formation of the functional structure of two RNA domains. These complexes affected not only elongation factor-binding site in the rRNA but also the functional structure of the decoding site. A gene coding for the Prp12 which interacts with Prp1 (pre-mRNA processing 1) of fission yeast which is a component of U4/U6 snRNP was cloned. As a result, it was elucidated that the interaction of U4/U6 snRNP with U2 snRNP which is necessary for conversion of a commitment complex to a spliceosome at the early stage of splicing depends on not only interaction due to hydrogen bonding between U2 and U6 snRNAs, but also interaction among proteins bound to the snRNAs.