Characterization of a family of RanBP2-type zinc fingers that can recognize single-stranded RNA.
The recognition of single-stranded RNA (ssRNA) is an important aspect of gene regulation, and a number of different classes of protein domains that recognize ssRNA in a sequence-specific manner have been identified. Recently, we demonstrated that the RanBP2-type zinc finger (ZnF) domains from the human splicing factor ZnF Ran binding domain-containing protein 2 (ZRANB2) can bind to a sequence containing the consensus AGGUAA. Six other human proteins, namely, Ewing's sarcoma (EWS), translocated in liposarcoma (TLS)/FUS, RNA-binding protein 56 (RBP56), RNA-binding motif 5 (RBM5), RNA-binding motif 10 (RBM10) and testis-expressed sequence 13A (TEX13A), each contains a single ZnF with homology to the ZRANB2 ZnFs, and several of these proteins have been implicated in the regulation of mRNA processing. Here, we show that all of these ZnFs are able to bind with micromolar affinities to ssRNA containing a GGU motif. NMR titration data reveal that binding is mediated by the corresponding surfaces on each ZnF, and we also show that sequence selectivity is largely limited to the GGU core motif and that substitution of the three flanking adenines that were selected in our original selection experiment has a minimal effect on binding affinity. These data establish a subset of RanBP2-type ZnFs as a new family of ssRNA-binding motifs.
School of Molecular Bioscience, University of Sydney, Sydney, NSW, Australia.
Cuong D
CD
Nguyen
Robyn E
RE
Mansfield
Wilfred
W
Leung
Paula M
PM
Vaz
Fionna E
FE
Loughlin
Richard P
RP
Grant
Joel P
JP
Mackay
D020816:Amino Acid Motifs / Q000235:genetics; D001483:Base Sequence; D006801:Humans; D008297:Male; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D028861:Nuclear Pore Complex Proteins / Q000235:genetics / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000235:genetics; D012313:RNA / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D016335:Zinc Fingers / Q000235:genetics*
D018832:Molecular Chaperones; D028861:Nuclear Pore Complex Proteins; D014157:Transcription Factors; C094493:ran-binding protein 2; D012313:RNA