STRUCTURE-FUNCTION ANALYSIS IN NUCLEAR RNASE-P RNA

Eukaryotic ribonuclease P (RNase P) enzymes require both RNA and prote in subunits for activity in vivo and in vitro. We have undertaken an a nalysis of the complex RNA subunit of the nuclear holoenzyme in an eff ort to understand its structure and its similarities to and difference s from the bacterial ribozymes. Phylogenetic analysis, structure-sensi tive RNA footprinting, and directed mutagenesis reveal conserved secon dary and tertiary structures with both strong similarities to the bact erial consensus and distinctive features. The effects of mutations in the most highly conserved positions are being used to dissect the func tions of individual subdomains.