Investigating the mechanism and machinery of RNA editing.
RNA editing in trypanosomes produces mature mRNAs by posttranscriptional guide RNA (gRNA)-directed uridylate insertion and deletion. This article describes methods for the study of RNA editing with an emphasis on an in vitro editing system that was used to explore the general mechanism of editing and that can be adapted for more in-depth studies of this intriguing and important process. Methods used to investigate the macromolecular complex that catalyzes RNA editing are also described. This complex is composed of multiple proteins and contains several catalytic activities. It is in the early stages of characterization. The methods described here are intended to assist in its further analysis.
Stuart
K
K
Seattle Biomedical Research Institute, Washington 98109-1651, USA. kstuart@u.washington.edu
Kable
M L
ML
Allen
T E
TE
Lawson
S
S
D000818:Animals; D001483:Base Sequence; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005821:Genetic Techniques; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D017393:RNA Editing; D017394:RNA, Guide, Kinetoplastida / Q000235:genetics* / Q000378:metabolism; D016053:RNA, Protozoan / Q000235:genetics* / Q000378:metabolism; D016601:RNA-Binding Proteins / Q000378:metabolism; D014346:Trypanosoma brucei brucei / Q000235:genetics*
D016053:RNA, Protozoan; D016601:RNA-Binding Proteins; D017394:RNA, Guide, Kinetoplastida