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  • Direct Molecular Evidence for an Ancient, Conserved Developmental Toolkit Controlling Posttranscriptional Gene Regulation in Land Plants
Title: Direct Molecular Evidence for an Ancient, Conserved Developmental Toolkit Controlling Posttranscriptional Gene Regulation in Land Plants
Abstract In plants, miRNA production is orchestrated by a suite of proteins that control transcription of the pri-miRNA gene, post-transcriptional processing and nuclear export of the mature miRNA. Post-transcriptional processing of miRNAs is controlled by a pair of physically interacting proteins, hyponastic leaves 1 (HYL1) and Dicer-like 1 (DCL1). However, the evolutionary history and structural basis of the HYL1–DCL1 interaction is unknown. Here we use ancestral sequence reconstruction and functional characterization of ancestral HYL1 in vitro and in Arabidopsis thaliana to better understand the origin and evolution of the HYL1–DCL1 interaction and its impact on miRNA production and plant development. We found the ancestral plant HYL1 evolved high affinity for both double-stranded RNA (dsRNA) and its DCL1 partner before the divergence of mosses from seed plants (∼500 Ma), and these high-affinity interactions remained largely conserved throughout plant evolutionary history. Structural modeling and molecular binding experiments suggest that the second of two dsRNA-binding motifs (DSRMs) in HYL1 may interact tightly with the first of two C-terminal DCL1 DSRMs to mediate the HYL1–DCL1 physical interaction necessary for efficient miRNA production. Transgenic expression of the nearly 200 Ma-old ancestral flowering-plant HYL1 in A. thaliana was sufficient to rescue many key aspects of plant development disrupted by HYL1− knockout and restored near-native miRNA production, suggesting that the functional partnership of HYL1–DCL1 originated very early in and was strongly conserved throughout the evolutionary history of terrestrial plants. Overall, our results are consistent with a model in which miRNA-based gene regulation evolved as part of a conserved plant “developmental toolkit.”  more » « less
Award ID(s):
1817942
PAR ID:
10351542
Author(s) / Creator(s):
; ; ;
Editor(s):
Norwick, Katja
Date Published:
Journal Name:
Molecular Biology and Evolution
Volume:
38
Issue:
11
ISSN:
0737-4038
Page Range / eLocation ID:
4765 to 4777
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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