6I3R

Structure, dynamics and roX2-lncRNA binding of tandem double-stranded RNA binding domains dsRBD1/2 of Drosophila helicase MLE

Summary for 6I3R

• Entry DOI: 10.2210/pdb6i3r/pdb
• NMR Information: BMRB: 34326
• Descriptor: Dosage compensation regulator (1 entity in total)
• Functional Keywords: helicase, dsrbd, rna binding protein
• Biological source: Drosophila melanogaster (Fruit fly)
• Total number of polymer chains: 1
• Total formula weight: 28589.94

Authors

Jagtap, P.K.A.,Buelow, S.v.,Masiewicz, P.,Simon, B.,Hennig, J. (deposition date: 2018-11-07, release date: 2019-02-20, Last modification date: 2024-06-19)

Primary citation

Ankush Jagtap, P.K.,Muller, M.,Masiewicz, P.,von Bulow, S.,Hollmann, N.M.,Chen, P.C.,Simon, B.,Thomae, A.W.,Becker, P.B.,Hennig, J.
Structure, dynamics and roX2-lncRNA binding of tandem double-stranded RNA binding domains dsRBD1,2 of Drosophila helicase Maleless.
Nucleic Acids Res., 47:4319-4333, 2019

PubMed Abstract: Maleless (MLE) is an evolutionary conserved member of the DExH family of helicases in Drosophila. Besides its function in RNA editing and presumably siRNA processing, MLE is best known for its role in remodelling non-coding roX RNA in the context of X chromosome dosage compensation in male flies. MLE and its human orthologue, DHX9 contain two tandem double-stranded RNA binding domains (dsRBDs) located at the N-terminal region. The two dsRBDs are essential for localization of MLE at the X-territory and it is presumed that this involves binding roX secondary structures. However, for dsRBD1 roX RNA binding has so far not been described. Here, we determined the solution NMR structure of dsRBD1 and dsRBD2 of MLE in tandem and investigated its role in double-stranded RNA (dsRNA) binding. Our NMR and SAXS data show that both dsRBDs act as independent structural modules in solution and are canonical, non-sequence-specific dsRBDs featuring non-canonical KKxAXK RNA binding motifs. NMR titrations combined with filter binding experiments and isothermal titration calorimetry (ITC) document the contribution of dsRBD1 to dsRNA binding in vitro. Curiously, dsRBD1 mutants in which dsRNA binding in vitro is strongly compromised do not affect roX2 RNA binding and MLE localization in cells. These data suggest alternative functions for dsRBD1 in vivo.

PubMed: 30805612
DOI: 10.1093/nar/gkz125

Experimental method

SOLUTION NMR