Skip to main content Accessibility help
×
Home

UV cross-link mapping of the substrate-binding site of an RNase P ribozyme to a target mRNA sequence

  • AHMED F. KILANI (a1) and FENYONG LIU (a1) (a2)

Abstract

RNase P ribozyme cleaves an RNA helix that resembles the acceptor stem and T-stem structure of its natural ptRNA substrate. When covalently linked with a guide sequence, the ribozyme can function as a sequence-specific endonuclease and cleave any target RNA sequences that base pair with the guide sequence. Using a site-directed ultraviolet (UV) cross-linking approach, we have mapped the regions of the ribozyme that are in close proximity to a substrate that contains the mRNA sequence encoding thymidine kinase of human herpes simplex virus 1. Our data suggest that the cleavage site of the mRNA substrate is positioned at the same regions of the ribozyme that bind to the cleavage site of a ptRNA. The mRNA-binding domains include regions that interact with the acceptor stem and T-stem and in addition, regions that are unique and not in close contact with a ptRNA. Identification of the mRNA-binding site provides a foundation to study how RNase P ribozymes achieve their sequence specificity and facilitates the development of gene-targeting ribozymes.

Copyright

Corresponding author

Reprint requests to: Dr. Fenyong Liu, School of Public Health, 140 Warren Hall, University of California, Berkeley, California 94720, USA; e-mail: liu_fy@uclink4.berkeley.edu.

Keywords

Related content

Powered by UNSILO

UV cross-link mapping of the substrate-binding site of an RNase P ribozyme to a target mRNA sequence

  • AHMED F. KILANI (a1) and FENYONG LIU (a1) (a2)

Metrics

Full text views

Total number of HTML views: 0
Total number of PDF views: 0 *
Loading metrics...

Abstract views

Total abstract views: 0 *
Loading metrics...

* Views captured on Cambridge Core between <date>. This data will be updated every 24 hours.

Usage data cannot currently be displayed.