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

2017; Impact Journals LLC; Volume: 8; Issue: 63 Linguagem: Inglês

10.18632/oncotarget.22540

1949-2553

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

RNA and protein synthesis mechanisms

// Lan Lan 1, * , Minli Xing 2, * , Justin T. Douglas 2 , Philip Gao 3 , Robert P. Hanzlik 4 and Liang Xu 1, 5 1 Departments of Molecular Biosciences, The University of Kansas, Lawrence, KS, USA 2 Bio-NMR Core Facility, The University of Kansas, Lawrence, KS, USA 3 Protein Production Group, NIH COBRE in Protein Structure and Function, The University of Kansas, Lawrence, KS, USA 4 Department of Medicinal Chemistry, The University of Kansas, Lawrence, KS, USA 5 Department 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: xul@ku.edu 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 ABSTRACT 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.