Identification of antisense RNA stem-loops that inhibit RNA-protein interactions using a bacterial reporter system.
Nucleic Acids Res
; 38(10): 3489-501, 2010 Jun.
Article
en En
| MEDLINE
| ID: mdl-20156995
Many well-characterized examples of antisense RNAs from prokaryotic systems involve hybridization of the looped regions of stem-loop RNAs, presumably due to the high thermodynamic stability of the resulting loop-loop and loop-linear interactions. In this study, the identification of RNA stem-loops that inhibit U1A protein binding to the hpII RNA through RNA-RNA interactions was attempted using a bacterial reporter system based on phage lambda N-mediated antitermination. As a result, loop sequences possessing 7-8 base complementarity to the 5' region of the boxA element important for functional antitermination complex formation, but not the U1 hpII loop, were identified. In vitro and in vivo mutational analysis strongly suggested that the selected loop sequences were binding to the boxA region, and that the structure of the antisense stem-loop was important for optimal inhibitory activity. Next, in an attempt to demonstrate the ability to inhibit the interaction between the U1A protein and the hpII RNA, the rational design of an RNA stem-loop that inhibits U1A-binding to a modified hpII was carried out. Moderate inhibitory activity was observed, showing that it is possible to design and select antisense RNA stem-loops that disrupt various types of RNA-protein interactions.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
ARN Nuclear Pequeño
/
ARN sin Sentido
/
Ribonucleoproteína Nuclear Pequeña U1
Tipo de estudio:
Diagnostic_studies
Idioma:
En
Revista:
Nucleic Acids Res
Año:
2010
Tipo del documento:
Article
País de afiliación:
Japón
Pais de publicación:
Reino Unido