Antisense probes containing 2 aminoadenosine allow efficient depletion of U5 snRNP from HeLa splicing extracts

G. LAMM; B. BLENCOWE; B. SPROAT; A. IRIBARREN; U. RYDER; A, LAMOND

NUCLEIC ACIDS RESEARCH

OXFORD UNIV PRESS

Lugar: Oxford; Año: 1991 vol. 19 p. 3193 - 3198

0305-1048

RNA duplexes containing the modified base 2-aminoadenine in place of adenine are stabilized through the formation of three hydrogen bonds in 2-amlno A-U base pairs. Antlsense 2´-0-alkyloligoribonucleotide probes incorporating 2-aminoadenoslne are thus able to efficiently affinity select RNP particles which are otherwise inaccessible. This has allowed the efficient and specific depletion of U5 snRNP from HeLa cell nuclear splicing extracts. U5 snRNP is shown to be essential for spliceosome assembly and for both steps of pre-mRNA splicing. The absence of U5 snRNP prevents the stable association of U4/U6 but not U1 and U2 snRNPs with pre-mRNA.