TY - JOUR
T1 - Inhibition of Pre-mRNA Splicing by a Synthetic Blom7α-Interacting Small RNA
AU - Löscher, Marlies
AU - Schosserer, Markus
AU - Dausse, Eric
AU - Lee, Kiseok
AU - Ajuh, Paul M.
AU - Grillari-Voglauer, Regina
AU - Lamond, Angus I.
AU - Toulmé, Jean Jacques
AU - Grillari, Johannes
PY - 2012/10/29
Y1 - 2012/10/29
N2 - Originally the novel protein Blom7α was identified as novel pre-mRNA splicing factor that interacts with SNEVPrp19/Pso4, an essential protein involved in extension of human endothelial cell life span, DNA damage repair, the ubiquitin-proteasome system, and pre-mRNA splicing. Blom7α belongs to the heteronuclear ribonucleoprotein K homology (KH) protein family, displaying 2 KH domains, a well conserved and widespread RNA-binding motif. In order to identify specific sequence binding motifs, we here used Systematic Evolution of Ligands by Exponential Enrichment (SELEX) with a synthetic RNA library. Besides sequence motifs like (U/A)1-4 C2-6 (U/A)1-5, we identified an AC-rich RNA-aptamer that we termed AK48 (Aptamer KH-binding 48), binding to Blom7α with high affinity. Addition of AK48 to pre-mRNA splicing reactions in vitro inhibited the formation of mature spliced mRNA and led to a slight accumulation of the H complex of the spliceosome. These results suggest that the RNA binding activity of Blom7α might be required for pre-mRNA splicing catalysis. The inhibition of in-vitro splicing by the small RNA AK48 indicates the potential use of small RNA molecules in targeting the spliceosome complex as a novel target for drug development.
AB - Originally the novel protein Blom7α was identified as novel pre-mRNA splicing factor that interacts with SNEVPrp19/Pso4, an essential protein involved in extension of human endothelial cell life span, DNA damage repair, the ubiquitin-proteasome system, and pre-mRNA splicing. Blom7α belongs to the heteronuclear ribonucleoprotein K homology (KH) protein family, displaying 2 KH domains, a well conserved and widespread RNA-binding motif. In order to identify specific sequence binding motifs, we here used Systematic Evolution of Ligands by Exponential Enrichment (SELEX) with a synthetic RNA library. Besides sequence motifs like (U/A)1-4 C2-6 (U/A)1-5, we identified an AC-rich RNA-aptamer that we termed AK48 (Aptamer KH-binding 48), binding to Blom7α with high affinity. Addition of AK48 to pre-mRNA splicing reactions in vitro inhibited the formation of mature spliced mRNA and led to a slight accumulation of the H complex of the spliceosome. These results suggest that the RNA binding activity of Blom7α might be required for pre-mRNA splicing catalysis. The inhibition of in-vitro splicing by the small RNA AK48 indicates the potential use of small RNA molecules in targeting the spliceosome complex as a novel target for drug development.
UR - http://www.scopus.com/inward/record.url?scp=84868091632&partnerID=8YFLogxK
U2 - 10.1371/journal.pone.0047497
DO - 10.1371/journal.pone.0047497
M3 - Article
C2 - 23144703
AN - SCOPUS:84868091632
VL - 7
JO - PLoS ONE
JF - PLoS ONE
SN - 1932-6203
IS - 10
M1 - e47497
ER -