Structural Basis for the Discriminative Recognition of N6-Methyladenosine RNA by the Human YT521-B Homology Domain Family of Proteins |
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| Authors: | Chao Xu ((许 超)) Ke Liu Hazem Ahmed Peter Loppnau Matthieu Schapira Jinrong Min |
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| Affiliation: | From the ‡Structural Genomics Consortium, University of Toronto, Toronto, Ontario M5G 1L7 and ;the §Department of Physiology, University of Toronto, Toronto, Ontario M5S 1A8 Canada |
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| Abstract: | N6-Methyladenosine (m6A) is the most abundant internal modification in RNA and is specifically recognized by YT521-B homology (YTH) domain-containing proteins. Recently we reported that YTHDC1 prefers guanosine and disfavors adenosine at the position preceding the m6A nucleotide in RNA and preferentially binds to the GG(m6A)C sequence. Now we systematically characterized the binding affinities of the YTH domains of three other human proteins and yeast YTH domain protein Pho92 and determined the crystal structures of the YTH domains of human YTHDF1 and yeast Pho92 in complex with a 5-mer m6A RNA, respectively. Our binding and structural data revealed that the YTH domain used a conserved aromatic cage to recognize m6A. Nevertheless, none of these YTH domains, except YTHDC1, display sequence selectivity at the position preceding the m6A modification. Structural comparison of these different YTH domains revealed that among those, only YTHDC1 harbors a distinctly selective binding pocket for the nucleotide preceding the m6A nucleotide. |
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| Keywords: | crystal structure isothermal titration calorimetry (ITC) RNA binding protein RNA methylation RNA-protein interaction N6-methyladenosine |
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