Inhibition of pre-mRNA splicing by 5-fluoro-, 5-chloro-, and 5-bromouridine.
Pre-mRNA transcripts of the human beta-globin gene containing 5-fluoro-, 5-chloro-, and 5-bromouridine were tested for splicing in vitro. Pre-mRNA containing 5-fluorouridine was spliced accurately and efficiently in the nuclear extract from HeLa cells, whereas 5-chloro-, and 5-bromouridine containing transcripts were not spliced. Analysis of the splicing reactions by electrophoresis on nondenaturing polyacrylamide gels showed that the latter two transcripts were unable to form active splicing complexes. Treatment of HeLa cell cultures with 5-fluorouridine decreased the splicing activity of the nuclear extracts in a dose- and time-dependent fashion. The decrease in splicing activity of these extracts appears to be due in part to a decreased level of U-2 small nuclear RNA and the corresponding ribonucleoprotein particle, U2-snRNP.
Sierakowska
H
H
Department of Pharmacology, University of North Carolina, Chapel Hill 27599.
Shukla
R R
RR
Dominski
Z
Z
Kole
R
R
D000998:Antiviral Agents / Q000494:pharmacology; D001976:Bromouracil / Q000031:analogs & derivatives; D004247:DNA / Q000235:genetics; D005091:Exons; D005467:Floxuridine / Q000494:pharmacology; D005796:Genes; D005816:Genetic Complementation Test; D005914:Globins / Q000235:genetics; D006367:HeLa Cells / Q000187:drug effects / Q000378:metabolism; D006801:Humans; D010957:Plasmids; D012322:RNA Precursors / Q000037:antagonists & inhibitors* / Q000187:drug effects / Q000235:genetics; D012326:RNA Splicing / Q000187:drug effects*; D012342:RNA, Small Nuclear / Q000187:drug effects / Q000378:metabolism; D013698:Templates, Genetic; D014158:Transcription, Genetic / Q000187:drug effects; D014529:Uridine / Q000378:metabolism / Q000494:pharmacology / Q000031:analogs & derivatives
D000998:Antiviral Agents; D012322:RNA Precursors; D012342:RNA, Small Nuclear; D005467:Floxuridine; C000455:5-chlorouridine; D001976:Bromouracil; C001943:5-fluorouridine; D005914:Globins; D004247:DNA; C006824:5-bromouridine; D014529:Uridine