'RNA walk' a novel approach to study RNA-RNA interactions between a small RNA and its target.

Abstract:

:In this study we describe a novel method to investigate the RNA-RNA interactions between a small RNA and its target that we termed 'RNA walk'. The method is based on UV-induced AMT cross-linking in vivo followed by affinity selection of the hybrid molecules and mapping the intermolecular adducts by RT-PCR or real-time PCR. Domains carrying the cross-linked adducts fail to efficiently amplify by PCR compared with non-cross-linked domains. This method was calibrated and used to study the interaction between a special tRNA-like molecule (sRNA-85) that is part of the trypanosome signal recognition particle (SRP) complex and the ribosome. Four contact sites between sRNA-85 and rRNA were identified by 'RNA walk' and were further fine-mapped by primer extension. Two of the contact sites are expected; one contact site mimics the interaction of the mammalian Alu domain of SRP with the ribosome and the other contact sites include a canonical tRNA interaction. The two other cross-linked sites could not be predicted. We propose that 'RNA walk, is a generic method to map target RNA small RNAs interactions in vivo.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Lustig Y,Wachtel C,Safro M,Liu L,Michaeli S

doi

10.1093/nar/gkp872

subject

Has Abstract

pub_date

2010-01-01 00:00:00

pages

e5

issue

1

eissn

0305-1048

issn

1362-4962

pii

gkp872

journal_volume

38

pub_type

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