Elucidation of the aberrant 3\u2032 splice site selection by cancer-associated mutations on the U2AF1

Hisashi Yoshida,Yuko Nariai,Takeshi Urano,Sam-Yong Park,Eiji Obayashi,Kanako Kuwasako,Gyosuke Sakashita,Yutaka Muto

doi:10.1038/s41467-020-18559-6

Hisashi Yoshida, Yuko Nariai + Show 6 more

Open Access

https://doi.org/10.1038/s41467-020-18559-6

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Abstract

The accurate exclusion of introns by RNA splicing is critical for the production of mature mRNA. U2AF1 binds specifically to the 3´ splice site, which includes an essential AG dinucleotide. Even a single amino acid mutation of U2AF1 can cause serious disease such as certain cancers or myelodysplastic syndromes. Here, we describe the first crystal structures of wild-type and pathogenic mutant U2AF1 complexed with target RNA, revealing the mechanism of 3´ splice site selection, and how aberrant splicing results from clinically important mutations. Unexpected features of this mechanism may assist the future development of new treatments against diseases caused by splicing errors.

Highlights

The accurate exclusion of introns by RNA splicing is critical for the production of mature mRNA
The amino acids involving in RNA binding and pathogenic hot spot, Ser[34] and Gln[157] of human U2AF1, are all conserved in the fission yeast U2AF1, shown in Supplementary Fig. 1
In our previous crystal structure of the U2AF1 complex without RNA, the zinc finger domains (ZFs) lie against the β-sheet of the U2AF homology motif (UHM) and contact each other, consistent with mutational experiments that suggested RNA might interact with both ZFs22

Summary

Introduction

The accurate exclusion of introns by RNA splicing is critical for the production of mature mRNA. We describe the first crystal structures of wild-type and pathogenic mutant U2AF1 complexed with target RNA, revealing the mechanism of 3 ́ splice site selection, and how aberrant splicing results from clinically important mutations. Unexpected features of this mechanism may assist the future development of new treatments against diseases caused by splicing errors. U2AF small subunit, U2AF1, binds to the 3′ side of the boundary sequence between the exon and intron, known as the 3′ splice site (3′SS, shown in Fig. 1a)[12,13].

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