TY - JOUR
T1 - A synthetic small molecule stalls pre-mRNA splicing by promoting an early-stage U2AF2-RNA complex
AU - Chatrikhi, Rakesh
AU - Feeney, Callen F.
AU - Pulvino, Mary J.
AU - Alachouzos, Georgios
AU - MacRae, Andrew J.
AU - Falls, Zackary
AU - Rai, Sumit
AU - Brennessel, William W.
AU - Jenkins, Jermaine L.
AU - Walter, Matthew J.
AU - Graubert, Timothy A.
AU - Samudrala, Ram
AU - Jurica, Melissa S.
AU - Frontier, Alison J.
AU - Kielkopf, Clara L.
N1 - Publisher Copyright:
© 2021 Elsevier Ltd
PY - 2021/8/19
Y1 - 2021/8/19
N2 - Dysregulated pre-mRNA splicing is an emerging Achilles heel of cancers and myelodysplasias. To expand the currently limited portfolio of small-molecule drug leads, we screened for chemical modulators of the U2AF complex, which nucleates spliceosome assembly and is mutated in myelodysplasias. A hit compound specifically enhances RNA binding by a U2AF2 subunit. Remarkably, the compound inhibits splicing of representative substrates and stalls spliceosome assembly at the stage of U2AF function. Computational docking, together with structure-guided mutagenesis, indicates that the compound bridges the tandem U2AF2 RNA recognition motifs via hydrophobic and electrostatic moieties. Cells expressing a cancer-associated U2AF1 mutant are preferentially killed by treatment with the compound. Altogether, our results highlight the potential of trapping early spliceosome assembly as an effective pharmacological means to manipulate pre-mRNA splicing. By extension, we suggest that stabilizing assembly intermediates may offer a useful approach for small-molecule inhibition of macromolecular machines.
AB - Dysregulated pre-mRNA splicing is an emerging Achilles heel of cancers and myelodysplasias. To expand the currently limited portfolio of small-molecule drug leads, we screened for chemical modulators of the U2AF complex, which nucleates spliceosome assembly and is mutated in myelodysplasias. A hit compound specifically enhances RNA binding by a U2AF2 subunit. Remarkably, the compound inhibits splicing of representative substrates and stalls spliceosome assembly at the stage of U2AF function. Computational docking, together with structure-guided mutagenesis, indicates that the compound bridges the tandem U2AF2 RNA recognition motifs via hydrophobic and electrostatic moieties. Cells expressing a cancer-associated U2AF1 mutant are preferentially killed by treatment with the compound. Altogether, our results highlight the potential of trapping early spliceosome assembly as an effective pharmacological means to manipulate pre-mRNA splicing. By extension, we suggest that stabilizing assembly intermediates may offer a useful approach for small-molecule inhibition of macromolecular machines.
KW - S34F mutant
KW - U2AF
KW - U2AF
KW - U2AF1
KW - myelodysplastic syndrome
KW - ribonucleoprotein targeting
KW - spliceosome inhibition
KW - splicing factor mutation
KW - therapeutic strategy
UR - http://www.scopus.com/inward/record.url?scp=85112540054&partnerID=8YFLogxK
U2 - 10.1016/j.chembiol.2021.02.007
DO - 10.1016/j.chembiol.2021.02.007
M3 - Article
C2 - 33689684
AN - SCOPUS:85112540054
SN - 2451-9456
VL - 28
SP - 1145-1157.e6
JO - Cell Chemical Biology
JF - Cell Chemical Biology
IS - 8
ER -