Analysis of RNA folding and ribonucleoprotein assembly by single-molecule fluorescence spectroscopy.
Methods Mol Biol
; 875: 271-95, 2012.
Article
em En
| MEDLINE
| ID: mdl-22573447
ABSTRACT
To execute their diverse range of biological functions, RNA molecules must fold into specific tertiary structures and/or associate with one or more proteins to form ribonucleoprotein (RNP) complexes. Single-molecule fluorescence spectroscopy is a powerful tool for the study of RNA folding and RNP assembly processes, directly revealing different conformational subpopulations that are hidden in conventional ensemble measurements. Moreover, kinetic processes can be observed without the need to synchronize a population of molecules. In this chapter, we describe the fluorescence spectroscopic methods used for single-molecule measurements of freely diffusing or immobilized RNA molecules or RNA-protein complexes. We also provide practical protocols to prepare the fluorescently labeled RNA and protein molecules required for such studies. Finally, we provide two examples of how these various preparative and spectroscopic methods are employed in the study of RNA folding and RNP assembly processes.
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Ribonucleoproteínas
/
Espectrometria de Fluorescência
/
Dobramento de RNA
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2012
Tipo de documento:
Article
País de afiliação:
Estados Unidos