Polyethylene glycol molecular crowders enhance the catalytic ability of bimolecular bacterial RNase P ribozymes

Abstract

The modular structure of bacterial ribonuclease P (RNase P) ribozymes, which recognize tertiary structures of precursor tRNAs (pre-tRNAs) to cleave their 5′ leader sequence, can be dissected physically into the two structured domain RNAs (S-domain and C-domain). Separately prepared S-domain RNA and C-domain RNA assemble to form bimolecular forms of RNase P ribozymes. We analyzed the effects of polyethylene glycols (PEGs) on pre-tRNA cleavage catalyzed by bimolecular RNase P ribozymes to examine the effects of molecular crowding on the reaction. PEG molecular crowders significantly enhanced the activities of bimolecular RNase P ribozymes, some of which were hardly active without PEGs.

Keywords:

• Modular structure
• molecular crowding
• ribozyme
• ribonuclease P
• RNA domain
• tRNA

Additional information

Funding

This work was supported by MEXT KAKENHI Grant Number JP15K05561 (to Y.I.). This work was also supported partly by University of Toyama Discretionary Funds of the President “Toyama RNA Research Alliance” (to Y.I. and S.M.).