Abstract
Ribonuclease P (RNase P) is an essential enzyme that processes the 5′ leader sequence of precursor tRNA. Eubacterial RNase P is an RNA enzyme, while its eukaryotic counterpart acts as catalytic ribonucleoprotein, consisting of RNA and numerous protein subunits. To study the latter form, we reconstitute human RNase P activity, demonstrating that the subunits H1 RNA, Rpp21, and Rpp29 are sufficient for 5′ cleavage of precursor tRNA. The reconstituted RNase P precisely delineates its cleavage sites in various substrates and hydrolyzes the phosphodiester bond. Rpp21 and Rpp29 facilitate catalysis by H1 RNA, which seems to require a phylogenetically conserved pseudoknot structure for function. Unexpectedly, Rpp29 forms a catalytic complex with M1 RNA of E. coli RNase P. The results uncover the core components of eukaryotic RNase P, reveal its evolutionary origin in translation, and provide a paradigm for studying RNA-based catalysis by other nuclear and nucleolar ribonucleoprotein enzymes.
| Original language | English |
|---|---|
| Pages (from-to) | 925-935 |
| Number of pages | 11 |
| Journal | Molecular Cell |
| Volume | 12 |
| Issue number | 4 |
| DOIs | |
| State | Published - Oct 2003 |
Bibliographical note
Funding Information:We thank Idit Goldshmidt and Amikam Cohen (Hebrew University) for providing us with yeast nuclear extracts and Cecilia Guerrier-Takada (Yale University) for advising us to set up the 2D-TLC. Y.B.-A. is supported by a postdoctoral fellowship from the Faculty of Medicine, the Hebrew University. This research is supported by the United States-Israel Binational Science Foundation (grant # 2001-017) and Israel Science Foundation (grant #549/01) to N.J. N.J. is the recipient of a Kahanoff Foundation Fellowship.