Biomolecular NMR Assignments: The 1H, 15N and 13C resonance assignments of the low-complexity domain from the oncogenic fusion protein EWS-FLI1 (Libich Lab)
Courtney N. Johnson,
Stephen P. Holloway &
David S. Libich
The RNA-binding protein EWS is a multifunctional protein with roles in the regulation of transcription and RNA splicing. It is one of the FET (FUS, EWS, and TAF15) family of RNA binding proteins that contain an intrinsically disordered, low-complexity N-terminal domain. The FET family proteins are prone to chromosomal translocations, often fusing their low-complexity domain with a transcription factor derived DNA-binding domain, that are oncogenic drivers in several leukemias and sarcomas. The fusion protein disrupts the normal function of cells through non-canonical DNA binding and alteration of normal transcriptional programs. However, the exact mechanism for how the intrinsically disordered domain contributes to aberrant DNA binding and abnormal transcription is unknown. The purification and 1H, 13C, and 15N backbone resonance assignments of the amino-terminal domain comprising 264 residues of EWS are described. This segment is common to all known EWS-fusions that are the hallmark of the pediatric cancer Ewing sarcoma. This domain is intrinsically disordered and features significant sequence degeneracy resulting in spectra with poor chemical shift dispersion. To alleviate this problem, the domain was divided into three overlapping fragments, reducing the complexity of the spectra and enabling an almost complete backbone resonance assignment of the full domain. These solution NMR chemical shift assignments represent the first steps towards understanding, at atomic resolution, how the low-complexity domain of EWS contributes to the aberrant functions of its oncogenic fusion proteins.
Since 2004, UT Health San Antonio, Greehey Children’s Cancer Research Institute’s (Greehey CCRI) mission has been to advance scientific knowledge relevant to childhood cancer, contribute to the understanding of its causes, and accelerate the translation of knowledge into novel therapies. Greehey CCRI strives to have a national and global impact on childhood cancer through the discovery, development, and dissemination of new scientific knowledge. Our mission consists of three key areas — research, clinical, and education.