Abstract
In higher eukaryotes, the dsRNA binding proteins (dsRBPs) assist the corresponding Dicer in the cleavage of dsRNA precursors to effect post-transcriptional gene regulation through RNA interference. In contrast, the DRB7.2:DRB4 complex in Arabidopsis thaliana acts as a potent inhibitor of Dicer-like 3 (DCL3) processing by sequestering endogenous inverted-repeat dsRNA precursors. DRB7.2 possesses a single dsRNA Binding Domain (dsRBD) flanked by unstructured N- and C-terminal regions. Whereas, DRB4 has two concatenated N-terminal dsRBDs and a long unstructured C-terminus harboring a small domain of unidentified function, D3. Here, we present near-complete backbone and partial side chain assignments of the interaction domains, DRB7.2M (i.e., DRB7.2 (71–162)) and DRB4D3 (i.e., DRB4 (294–355)) as a complex. Our findings establish the groundwork for future structural, dynamic, and functional research on DRB7.2 and DRB4, and provide clues for the endo-IR pathway in plants.
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Data Availability
Under accession number BMRB-51790 for DRB7.2M and accession number BMRB-51791 for DRB4D3, the 1H, 13C, and 15N chemical shifts of the interacting domains have been deposited to the BioMagResBank (www.bmrb.wisc.edu).
Change history
01 November 2023
A Correction to this paper has been published: https://doi.org/10.1007/s12104-023-10152-4
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Acknowledgements
We acknowledge Prof. Patrice Dunoyer (Institut de Biologie Moléculaire des Plantes du CNRS, Strasbourg) for the generous gift of recombinant DRB7.2 plasmid. Authors acknowledge Protein Expression and Purification Core Facility, EMBL Heidelberg for the generous gift of the pETtrx_1b vector.
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This work was supported by the Council of Scientific and Industrial Research, Government of India through a CSIR-FIRST grant (MLP161). SP acknowledges research fellowships from the Department of Biotechnology (DBT) and the Council of Scientific and Industrial Research (CSIR).
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MVD planned the study and designed experiments. SP and MVD performed experiments and interpreted the results. MVD and SP wrote the manuscript.
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Paturi, S., Deshmukh, M.V. NMR resonance assignments of 18.5 kDa complex of Arabidopsis thaliana DRB7.2:DRB4 interaction domains. Biomol NMR Assign 17, 173–178 (2023). https://doi.org/10.1007/s12104-023-10137-3
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DOI: https://doi.org/10.1007/s12104-023-10137-3