Abstract
NCYM is a cis-antisense gene of MYCN oncogene and encodes an oncogenic protein that stabilizes MYCN via inhibition of GSK3b. High NCYM expression levels are associated with poor clinical outcomes in human neuroblastomas, and NCYM overexpression promotes distant metastasis in animal models of neuroblastoma. Using vacuum-ultraviolet circular dichroism and small-angle X-ray scattering, we previously showed that NCYM has high flexibility with partially folded structures; however, further structural characterization is required for the design of anti-cancer agents targeting NCYM. Here we report the 1H, 15N and 13C nuclear magnetic resonance assignments of NCYM. Secondary structure prediction using Secondary Chemical Shifts and TALOS-N analysis demonstrates that the structure of NCYM is essentially disordered, even though residues in the central region of the peptide clearly present a propensity to adopt a dynamic helical structure. This preliminary study provides foundations for further analysis of interaction between NCYM and potential partners.
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Funding
This project was supported by French Infrastructure for Integrated Structural Biology (FRISBI) Grant No. ANR-10-INSB-05 (CR), and by the AMED eASIA JRP (22jm0210092h0002) (YH, YS, TT). This work benefited from the Japanese-French bilateral research cooperation program “Sakura” 2023–2024 (49615UB) (JP), and JSPS Bilateral Program (JPJSBP120233212) (TM), and from the Mission pour les Initiatives Transverses et Interdisciplinaires (CNRS) (LifeAdapt research and Bioquant projects) (JP). J.P. is grateful to the Institut Universitaire de France for providing additional time to be dedicated to research.
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KN, FK and YH sub-cloned, expressed and purified the recombinant protein. AM recorded and analyzed the NMR spectra with PB and CR. She also prepared the figures and contributed to the writing. AM, CR and YS wrote the manuscript. JP, TM, YS and TT found the funding and coordinated the project with CR. All authors reviewed the manuscript.
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Mouhand, A., Nakatani, K., Kono, F. et al. 1H, 13C and 15N backbone and side-chain resonance assignments of the human oncogenic protein NCYM. Biomol NMR Assign (2024). https://doi.org/10.1007/s12104-024-10169-3
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DOI: https://doi.org/10.1007/s12104-024-10169-3