Abstract
FUS, a nuclear RNA-binding protein, plays multiple roles in RNA processing. Five specific FUS-binding RNA sequence/structure motifs have been proposed, but their affinities for FUS have not been directly compared. Here we find that human FUS binds all these sequences with Kdapp values spanning a 10-fold range. Furthermore, some RNAs that do not contain any of these motifs bind FUS with similar affinity. FUS binds RNA in a length-dependent manner, consistent with a substantial non-specific component to binding. Finally, investigation of FUS binding to different nucleic acids shows that it binds single-stranded DNA with three-fold lower affinity than ssRNA of the same length and sequence, while binding to double-stranded nucleic acids is weaker. We conclude that FUS has quite general nucleic acid-binding activity, with the various proposed RNA motifs being neither necessary for FUS binding nor sufficient to explain its diverse binding partners.
Highlights
FUsed in Sarcoma (FUS, known as Translocated in LipoSarcoma, TLS), is an abundant nuclear protein that has been implicated in transcription, mRNA splicing and mRNA transport [1,2,3]
To characterize the features of RNA targets necessary for FUS binding, we have thoroughly evaluated the binding affinities of FUS with all five published RNA motifs and additional sequences, using electrophoretic mobility shift assays (EMSAs)
We measured the binding of E. coli-expressed FUS protein to eight RNAs including the five published motifs and three negative control sequences (Supplementary Table S1)
Summary
FUsed in Sarcoma (FUS, known as Translocated in LipoSarcoma, TLS), is an abundant nuclear protein that has been implicated in transcription, mRNA splicing and mRNA transport [1,2,3]. Mutations in FUS are detected in ∼5% of familial ALS (amyotrophic lateral sclerosis) patients as well as in sporadic ALS [4,5]. ALS is a progressive motor neuron disease characterized by loss of the upper and lower motor neurons [6]. Patients typically die within 3–5 years after onset of the disease. Dysregulation of RNA is emerging as a pathogenic mechanism in ALS. Understanding the biology and biochemistry of the FUS protein may provide insights into how this protein can potentially cause the onset of the disease
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