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

• Published in: Nucleic acids research Vol. 19; no. 12; pp. 3193 - 3198
• Main Authors: LAMM, G. M, BLENCOWE, B. J, SPROAT, B. S, IRIBARREN, A. M, RYDER, U, LAMOND, A. I
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 25.06.1991
• Subjects: 2-aminoadenosine; Adenosine - analogs & derivatives; Adenosine - chemistry; Base Sequence; Biological and medical sciences; DNA; Electrophoresis, Polyacrylamide Gel; Endoribonucleases - metabolism; Fundamental and applied biological sciences. Psychology; HeLa Cells; Humans; Hydrogen Bonding; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; Nucleic Acid Conformation; Ribonuclease H; ribonucleoproteins; Ribonucleoproteins - metabolism; Ribonucleoproteins - physiology; Ribonucleoproteins, Small Nuclear; RNA Precursors - metabolism; RNA Probes - chemistry; RNA Probes - metabolism; RNA Splicing - physiology; RNA, Antisense - chemistry; RNA, Antisense - metabolism; Transcription. Transcription factor. Splicing. Rna processing; Human; Purine nucleotide; Spliceosome; Ribonucleoprotein; Cell line; Messenger RNA; Splicing; Molecular assembly; Antisense RNA; Molecular probe
• ISSN: 0305-1048; 1362-4962
• DOI: 10.1093/nar/19.12.3193

RNA duplexes containing the modified base 2-amino-adenine in place of adenine are stabilized through the formation of three hydrogen bonds in 2-amino A.U base pairs. Antisense 2'-O-alkyloligoribonucleotide probes incorporating 2-aminoadenosine 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.