Biochemical and biophysical characterization of the nucleic acid binding properties of the RNA/DNA binding protein EWS.
EWS
G-quadruplex
NMR
R-loop
intrinsically disordered protein
Journal
Biopolymers
ISSN: 1097-0282
Titre abrégé: Biopolymers
Pays: United States
ID NLM: 0372525
Informations de publication
Date de publication:
May 2023
May 2023
Historique:
revised:
06
03
2023
received:
13
12
2022
accepted:
08
03
2023
pmc-release:
01
05
2024
medline:
26
5
2023
pubmed:
18
3
2023
entrez:
17
3
2023
Statut:
ppublish
Résumé
EWS is a member of the FET family of RNA/DNA binding proteins that regulate crucial phases of nucleic acid metabolism. EWS comprises an N-terminal low-complexity domain (LCD) and a C-terminal RNA-binding domain (RBD). The RBD is further divided into three RG-rich regions, which flank an RNA-recognition motif (RRM) and a zinc finger (ZnF) domain. Recently, EWS was shown to regulate R-loops in Ewing sarcoma, a pediatric bone and soft-tissue cancer in which a chromosomal translocation fuses the N-terminal LCD of EWS to the C-terminal DNA binding domain of the transcription factor FLI1. Though EWS was shown to directly bind R-loops, the binding mechanism was not elucidated. In the current study, the RBD of EWS was divided into several constructs, which were subsequently assayed for binding to various nucleic acid structures expected to form at R-loops, including RNA stem-loops, DNA G-quadruplexes, and RNA:DNA hybrids. EWS interacted with all three nucleic acid structures with varying affinities and multiple domains contributed to binding each substrate. The RRM and RG2 region appear to bind nucleic acids promiscuously while the ZnF displayed more selectivity for single-stranded structures. With these results, the structural underpinnings of EWS recognition and binding of R-loops and other nucleic acid structures is better understood.
Identifiants
pubmed: 36929870
doi: 10.1002/bip.23536
pmc: PMC10233817
mid: NIHMS1899836
doi:
Substances chimiques
RNA-Binding Protein EWS
0
RNA-Binding Proteins
0
DNA-Binding Proteins
0
DNA
9007-49-2
RNA
63231-63-0
Nucleic Acids
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
e23536Subventions
Organisme : NIGMS NIH HHS
ID : R01 GM140127
Pays : United States
Organisme : NIGMS NIH HHS
ID : GM140127
Pays : United States
Informations de copyright
© 2023 Wiley Periodicals LLC.
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