TY - JOUR
T1 - Sequence-specific RNA recognition by an RGG motif connects U1 and U2 snRNP for spliceosome assembly
AU - De Vries, Tebbe
AU - Martelly, William
AU - Campagne, Sébastien
AU - Sabath, Kevin
AU - Sarnowski, Chris P.
AU - Wong, Jason
AU - Leitner, Alexander
AU - Jonas, Stefanie
AU - Sharma, Shalini
AU - Allain, Frédéric H.T.
N1 - Funding Information:
Light Source PXI and PXIII beamlines, in particular Vincent Olieric (PSI, Switzerland) for support during data collection and Nenad Ban for access to crystallization equipment. This work was supported by ETH Zu€rich (Research Grant ETH-24 16-2 to A.L., F.H.-T.A.), the Swiss National Science Foundation through
Publisher Copyright:
© 2022 National Academy of Sciences. All rights reserved.
PY - 2022/2/8
Y1 - 2022/2/8
N2 - In mammals, the structural basis for the interaction between U1 and U2 small nuclear ribonucleoproteins (snRNPs) during the early steps of splicing is still elusive. The binding of the ubiquitin-like (UBL) domain of SF3A1 to the stem-loop 4 of U1 snRNP (U1-SL4) contributes to this interaction. Here, we determined the 3D structure of the complex between the UBL of SF3A1 and U1-SL4 RNA. Our crystallography, NMR spectroscopy, and cross-linking mass spectrometry data show that SF3A1-UBL recognizes, sequence specifically, the GCG/CGC RNA stem and the apical UUCG tetraloop of U1-SL4. In vitro and in vivo mutational analyses support the observed intermolecular contacts and demonstrate that the carboxyl-terminal arginine-glycine-glycine-arginine (RGGR) motif of SF3A1-UBL binds sequence specifically by inserting into the RNA major groove. Thus, the characterization of the SF3A1-UBL/U1-SL4 complex expands the repertoire of RNA binding domains and reveals the capacity of RGG/RG motifs to bind RNA in a sequencespecific manner.
AB - In mammals, the structural basis for the interaction between U1 and U2 small nuclear ribonucleoproteins (snRNPs) during the early steps of splicing is still elusive. The binding of the ubiquitin-like (UBL) domain of SF3A1 to the stem-loop 4 of U1 snRNP (U1-SL4) contributes to this interaction. Here, we determined the 3D structure of the complex between the UBL of SF3A1 and U1-SL4 RNA. Our crystallography, NMR spectroscopy, and cross-linking mass spectrometry data show that SF3A1-UBL recognizes, sequence specifically, the GCG/CGC RNA stem and the apical UUCG tetraloop of U1-SL4. In vitro and in vivo mutational analyses support the observed intermolecular contacts and demonstrate that the carboxyl-terminal arginine-glycine-glycine-arginine (RGGR) motif of SF3A1-UBL binds sequence specifically by inserting into the RNA major groove. Thus, the characterization of the SF3A1-UBL/U1-SL4 complex expands the repertoire of RNA binding domains and reveals the capacity of RGG/RG motifs to bind RNA in a sequencespecific manner.
KW - RGG motif
KW - Spliceosome assembly
KW - Splicing
KW - Ubiquitin-like domain
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U2 - 10.1073/pnas.2114092119
DO - 10.1073/pnas.2114092119
M3 - Article
C2 - 35101980
AN - SCOPUS:85123986930
SN - 0027-8424
VL - 119
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 6
M1 - e2114092119
ER -