RNA splicing products formed with isolated fractions from HeLa cells are associated with fast-sedimenting complexes
Three fractions (designated Ia, Ib, and II) have been isolated from HeLa cell nuclear extracts that are required for splicing of adenovirus and human US -globin RNA transcripts in vitro. The incubation of two of the fractions (Ib and II) in the presence of ATP resulted in cleavage of precursor mRNA at the 5' splice site and formation of the intron-exon lariat. Addition of fraction Ia to the combination of Ib and II resulted in the formation of spliced RNA and the intron lariat TSP-labelled 40s ribosomes were utilized. When fraction II was incubated with precursor RNA in the presence of ATP and the resulting products were sedimented through sucrose gradients, a 30S complex was detected that contained precursor RNA. The combination of fractions Ib and II resulted in the production of a 55S complex that contained the 5' exon as a prominent RNA species. The combination of fractions I (containing Ia and Ib) and II resulted in the formation of the 55S complex and material sedimenting between 40 S and 20 S, in which the predominant RNA species was spliced RNA.
- Research Organization:
- Sloan-Kettering Institute for Cancer Research, New York, NY
- OSTI ID:
- 6358580
- Journal Information:
- Proc. Natl. Acad. Sci. U.S.A.; (United States), Vol. 83:4
- Country of Publication:
- United States
- Language:
- English
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MESSENGER-RNA
MOLECULAR STRUCTURE
PHOSPHORUS 32
TRACER TECHNIQUES
ADENOVIRUS
ATP
GLOBIN
HELA CELLS
IN VITRO
MAN
RIBOSOMES
SEDIMENTATION
TRANSCRIPTION
ANIMALS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CELL CONSTITUENTS
DAYS LIVING RADIOISOTOPES
ISOTOPE APPLICATIONS
ISOTOPES
LIGHT NUCLEI
MAMMALS
MICROORGANISMS
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDES
ODD-ODD NUCLEI
ONCOGENIC VIRUSES
ORGANIC COMPOUNDS
ORGANOIDS
PARASITES
PHOSPHORUS ISOTOPES
PRIMATES
PROTEINS
RADIOISOTOPES
RNA
VERTEBRATES
VIRUSES
550201* - Biochemistry- Tracer Techniques