Research Papers:

Human oncoprotein Musashi-2 N-terminal RNA recognition motif backbone assignment and identification of RNA-binding pocket

Lan Lan, Minli Xing, Justin T. Douglas, Philip Gao, Robert P. Hanzlik and Liang Xu _

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Oncotarget. 2017; 8:106587-106597. https://doi.org/10.18632/oncotarget.22540

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Lan Lan1,*, Minli Xing2,*, Justin T. Douglas2, Philip Gao3, Robert P. Hanzlik4 and Liang Xu1,5

1Departments of Molecular Biosciences, The University of Kansas, Lawrence, KS, USA

2Bio-NMR Core Facility, The University of Kansas, Lawrence, KS, USA

3Protein Production Group, NIH COBRE in Protein Structure and Function, The University of Kansas, Lawrence, KS, USA

4Department of Medicinal Chemistry, The University of Kansas, Lawrence, KS, USA

5Department of Radiation Oncology, The University of Kansas Cancer Center, Kansas City, KS, USA

*These authors contributed equally to this work

Correspondence to:

Liang Xu, email: [email protected]

Keywords: RNA-binding protein; RNA-binding pocket; nuclear magnetic resonance; backbone assignment; Musashi

Received: September 07, 2017     Accepted: October 30, 2017     Published: November 20, 2017


RNA-binding protein Musashi-2 (MSI2) is a key regulator in stem cells, it is over-expressed in a variety of cancers and its higher expression is associated with poor prognosis. Like Musashi-1, it contains two N-terminal RRMs (RNA-recognition Motifs, also called RBDs (RNA-binding Domains)), RRM1 and RRM2, which mediate the binding to their target mRNAs. Previous studies have obtained the three-dimensional structures of the RBDs of Musashi-1 and the RBD1:RNA complex. Here we show the binding of MSI2-RRM1 to a 15nt Numb RNA in Fluorescence Polarization assay and time resolved Fluorescence Resonance Energy Transfer assay. Using nuclear magnetic resonance (NMR) spectroscopy we assigned the backbone resonances of MSI2-RRM1, and characterized the direct interaction of RRM1 to Numb RNA r(GUAGU). Our NMR titration and structure modeling studies showed that MSI2-RRM1 and MSI1-RBD1 have similar RNA binding events and binding pockets. This work adds significant information to MSI2-RRM1 structure and RNA binding pocket, and contributes to the development of MSI2 specific and MSI1/MSI2 dual inhibitors.

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