TY - JOUR
T1 - Molecular principles underlying dual RNA specificity in the Drosophila SNF protein
AU - Weber, Gert
AU - Dekoster, Gregory T.
AU - Holton, Nicole
AU - Hall, Kathleen B.
AU - Wahl, Markus C.
N1 - Funding Information:
This work was supported in part by grant 1R01 GM096444 from NIH (to K.B.H.) and grant SFB740-3 from the Deutsche Forschungsgemeinschaft (to M.C.W.). RNAs for crystallization were a gift from Agilent labs (Dr. Doug Dellinger). RNAs for NMR were prepared by Michael Rau. We acknowledge access to beamlines BL14.1 and 14.2 of the BESSY II storage ring (Berlin, Germany) via the Joint Berlin MX-Laboratory, funded by the Helmholtz-Zentrum Berlin fur Materialien und Energie, the Freie Universität Berlin, the Humboldt-Universität zu Berlin, the Max-Delbruck Centrum für Molekulare Medizin and the Leibniz-Institut für Molekulare Pharmakologie.
Funding Information:
This work was supported in part by grant 1R01 GM096444 from NIH (to K.B.H.) and grant SFB740-3 from the Deutsche Forschungsgemeinschaft (to M.C.W.). RNAs for crystallization were a gift from Agilent labs (Dr. Doug Dellinger). RNAs for NMR were prepared by Michael Rau. We acknowledge access to beamlines BL14.1 and 14.2 of the BESSY II storage ring (Berlin, Germany) via the Joint Berlin MX-Laboratory, funded by the Helmholtz-Zentrum Berlin für Materialien und Energie, the Freie Universität Berlin, the Humboldt-Universität zu Berlin, the Max-Delbrück Centrum für Molekulare Med-izin and the Leibniz-Institut für Molekulare Pharmakologie.
Publisher Copyright:
© 2018 The Author(s).
PY - 2018/12/1
Y1 - 2018/12/1
N2 - The first RNA recognition motif of the Drosophila SNF protein is an example of an RNA binding protein with multi-specificity. It binds different RNA hairpin loops in spliceosomal U1 or U2 small nuclear RNAs, and only in the latter case requires the auxiliary U2A′ protein. Here we investigate its functions by crystal structures of SNF alone and bound to U1 stem-loop II, U2A′ or U2 stem-loop IV and U2A′, SNF dynamics from NMR spectroscopy, and structure-guided mutagenesis in binding studies. We find that different loop-closing base pairs and a nucleotide exchange at the tips of the loops contribute to differential SNF affinity for the RNAs. U2A′ immobilizes SNF and RNA residues to restore U2 stem-loop IV binding affinity, while U1 stem-loop II binding does not require such adjustments. Our findings show how U2A′ can modulate RNA specificity of SNF without changing SNF conformation or relying on direct RNA contacts.
AB - The first RNA recognition motif of the Drosophila SNF protein is an example of an RNA binding protein with multi-specificity. It binds different RNA hairpin loops in spliceosomal U1 or U2 small nuclear RNAs, and only in the latter case requires the auxiliary U2A′ protein. Here we investigate its functions by crystal structures of SNF alone and bound to U1 stem-loop II, U2A′ or U2 stem-loop IV and U2A′, SNF dynamics from NMR spectroscopy, and structure-guided mutagenesis in binding studies. We find that different loop-closing base pairs and a nucleotide exchange at the tips of the loops contribute to differential SNF affinity for the RNAs. U2A′ immobilizes SNF and RNA residues to restore U2 stem-loop IV binding affinity, while U1 stem-loop II binding does not require such adjustments. Our findings show how U2A′ can modulate RNA specificity of SNF without changing SNF conformation or relying on direct RNA contacts.
UR - http://www.scopus.com/inward/record.url?scp=85048248691&partnerID=8YFLogxK
U2 - 10.1038/s41467-018-04561-6
DO - 10.1038/s41467-018-04561-6
M3 - Article
C2 - 29880797
AN - SCOPUS:85048248691
SN - 2041-1723
VL - 9
JO - Nature communications
JF - Nature communications
IS - 1
M1 - 2220
ER -