|
| Status |
Public on Oct 14, 2021 |
| Title |
Mechanism of siRNA production by a plant Dicer-RNA complex in dicing-competent conformation |
| Organism |
Arabidopsis thaliana |
| Experiment type |
Non-coding RNA profiling by high throughput sequencing
|
| Summary |
In eukaryotes, small RNAs (sRNAs) play critical roles in multiple biological processes. Dicer endonucleases are central to sRNA biogenesis. In plants, DICER-LIKE PROTEIN 3 (DCL3) produces 24-nt small interfering RNAs (siRNAs) that determine the specificity of the RNA-directed DNA methylation (RdDM) pathway. Here, we determined structure of a DCL3-pre-siRNA complex in an active dicing-competent state. The 5′-phosphorylated-A1 of the guide strand and the 1-nt 3′-overhang of the complementary strand are specifically recognized by a positively charged pocket and an aromatic cap, respectively. The 24-nt siRNA length dependence relies on the separation between the 5′-phosphorylated-end of the guide RNA and dual cleavage sites formed by the paired RNaseIII domains. These structural studies, complemented by functional data, reveal insights into the dicing principle for Dicers in general.
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| |
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| Overall design |
This dataset includes small RNA-seq to understand the in vivo molecular mechanism of Arabidopsis DCL3 protein.
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| Contributor(s) |
Xue Y, Zhong Z |
| Citation(s) |
34648373 |
| Submission date |
Jul 06, 2021 |
| Last update date |
Oct 16, 2021 |
| Contact name |
Zhenhui Zhong |
| E-mail(s) |
zhenhuizhong@gmail.com
|
| Organization name |
University of California, Los Angeles
|
| Department |
Department of Molecular, Cell and Developmental Biology
|
| Lab |
Jacobsen Lab
|
| Street address |
610 Charles E Young Dr East
|
| City |
Los Angeles |
| ZIP/Postal code |
90095 |
| Country |
USA |
| |
|
| Platforms (1) |
| GPL26208 |
Illumina NovaSeq 6000 (Arabidopsis thaliana) |
|
| Samples (20)
|
| GSM5608777 |
small RNA data of dcl234_rep2 |
| GSM5608778 |
small RNA data of DCL3-WT-1 in dcl234 |
| GSM5608779 |
small RNA data of DCL3-WT-3 in dcl234 |
| GSM5608780 |
small RNA data of DCL3 deletion 865-895 rep1 in dcl234 |
| GSM5608781 |
small RNA data of DCL3 deletion 865-895 rep2 in dcl234 |
| GSM5608782 |
small RNA data of DCL3 deletion 1356-1570 rep1 in dcl234 |
| GSM5608783 |
small RNA data of DCL3 deletion 1356-1570 rep2 in dcl234 |
| GSM5608784 |
small RNA data of DCL3-deletion 1450-1570 rep1 in dcl234 |
| GSM5608785 |
small RNA data of DCL3-deletion 1450-1570 rep2 in dcl234 |
| GSM5608786 |
small RNA data of DCL3 D1316A/E1319A/E1224A/D1228A rep1 in dcl234 |
| GSM5608787 |
small RNA data of DCL3 D1316A/E1319A/E1224A/D1228A rep2 in dcl234 |
| GSM5608788 |
small RNA data of DCL3 D1133A/E1136A/E1015A/D1019A rep1 in dcl234 |
| GSM5608789 |
small RNA data of DCL3 D1133A/E1136A/E1015A/D1019A rep2 in dcl234 |
| GSM5608790 |
small RNA data of DCL3 K695A/K957A/K903A rep1 in dcl234 |
| GSM5608791 |
small RNA data of DCL3 K695A/K957A/K903A rep2 in dcl234 |
| GSM5608792 |
small RNA data of DCL3 H909A rep1 in dcl234 |
| GSM5608793 |
small RNA data of DCL3 H909A rep2 in dcl234 |
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| Relations |
| BioProject |
PRJNA744279 |
| SRA |
SRP327236 |
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