Pre-mRNA with a trimethylguanosine cap structure can be spliced efficiently in vitro.
A chemically synthesized 2,2,7-trimethylguanosine cap (TMG-cap) analogue was added to an in vitro pre-mRNA splicing system. A slight inhibition of the splicing was observed, but it was much less remarkable than that by a 7-methylguanosine cap analogue. This result does not favor a direct role of the TMG-cap structure of small nuclear RNAs in pre-mRNA splicing, and suggests that the TMG-cap does not effectively interact with the 7-methylguanosine cap binding factors. Using the SP6 in vitro transcription system and a TMG-cap analogue, we prepared a beta-globin transcript which has a TMG-cap at the 5' end. This TMG-capped transcript was spliced accurately and efficiently in vitro.
Tokumoto
Y
Y
Department of Biology, Faculty of Science, Kyushu University, Fukuoka, Japan.
Iwase
R
R
Sekine
M
M
Hata
T
T
Ohshima
Y
Y
D001667:Binding, Competitive; D002467:Cell Nucleus / Q000502:physiology; D006367:HeLa Cells; D066298:In Vitro Techniques; D007700:Kinetics; D012315:RNA Caps; D012326:RNA Splicing; D013329:Structure-Activity Relationship
D012315:RNA Caps