Inhibition of splicing but not cleavage at the 5' splice site by truncating human US -globin pre-mRNA
Human US -globin mRNAs truncated in the second exon or in the first intron have been processed in vitro in a HeLa cell nuclear extract. Transcripts containing a fragment of the second exon as short as 53 nucleotides are efficiently spliced, whereas transcripts truncated 24 or 14 nucleotides downstream from the 3' splice site are spliced inefficiently, if at all. All of these transcripts, however, are efficiently and accurately cleaved at the 5' splice site. In contrast, RNA truncated in the first intron, 54 nucleotides upstream from the 3' splice site, is not processed at all (TSP)RNA products from the in vitro processing reactions were electrophoresed and autoradiographed. These findings suggest that cleavage at the 5' splice site and subsequent splicing steps - i.e., cleavage at the 3' splice site and exon ligation-need not be coupled. Anti-Sm serum inhibits the complete splicing reaction and cleavage at the 5' splice site, suggesting involvement of certain ribonucleoprotein particles in the cleavage reaction. ATP and MgS are required for cleavage at the 5' splice site at concentrations similar to those for the complete splicing reaction.
- Research Organization:
- Univ. of North Carolina, Chapel Hill, NC (United States)
- OSTI ID:
- 6293997
- 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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Related Subjects
MESSENGER-RNA
AUTORADIOGRAPHY
ENZYMATIC HYDROLYSIS
ATP
ELECTROPHORESIS
GLOBIN
HELA CELLS
IMMUNE SERUMS
IN VITRO
MAGNESIUM CHLORIDES
MOLECULAR STRUCTURE
NUCLEOPROTEINS
PHOSPHORUS 32
TRANSCRIPTION
ALKALINE EARTH METAL COMPOUNDS
BETA DECAY RADIOISOTOPES
BETA-MINUS DECAY RADIOISOTOPES
CHEMICAL REACTIONS
CHLORIDES
CHLORINE COMPOUNDS
DAYS LIVING RADIOISOTOPES
DECOMPOSITION
HALIDES
HALOGEN COMPOUNDS
HYDROLYSIS
ISOTOPES
LIGHT NUCLEI
LYSIS
MAGNESIUM COMPOUNDS
NUCLEI
NUCLEIC ACIDS
NUCLEOTIDES
ODD-ODD NUCLEI
ORGANIC COMPOUNDS
PHOSPHORUS ISOTOPES
PROTEINS
RADIOISOTOPES
RNA
SOLVOLYSIS
550601* - Medicine- Unsealed Radionuclides in Diagnostics