Chimeric DCL1-Partnering Proteins Provide Insights into the MicroRNA Pathway

Frontiers in Plant Science
Rodrigo S ReisPeter M Waterhouse

Abstract

In Arabidopsis thaliana, efficient microRNA (miRNA) production requires DICER-LIKE1 (DCL1) with the assistance of a partnering protein, DOUBLE-STRANDED RNA BINDING1 (DRB1) or DRB2. The presence of either of these DRB proteins is crucial to determine the mode of action of a miRNA; i.e., cleavage or translation inhibition. Here we studied the structural determinants for the role of DRB1 and DRB2 in the miRNA pathway. We developed a series of chimeric vectors encoding different functional domains of DRB1 and DRB2, and expressed these in the drb1 mutant background in Arabidopsis under the control of the native DRB1 promoter. Complementation of the drb1 developmental phenotype was used to assess the biological role that each functional domain of DRB1 and DRB2 mediates in the miRNA-guided transcript cleavage pathway. The DRB1 amino acid sequence differs considerably to that of DRB2, and analysis of drb1 transgenic lines revealed that the first dsRNA-binding domains of DRB1 and DRB2 are functionally similar; in contrast, the dsRBD2 of DRB1 and DRB2 appear functionally distinct. Our bioinformatic analysis further suggests that the C-terminal domain of DRB2 mediates a functional role in the miRNA pathway, whereas its counterpart in DRB1...Continue Reading

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Citations

Mar 28, 2020·Journal of Virology·Károly FátyolMárta Ludman
Sep 13, 2020·International Journal of Molecular Sciences·Yujie YanWentao Hu
May 25, 2021·Frontiers in Molecular Biosciences·Sneha Paturi, Mandar V Deshmukh

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Methods Mentioned

BETA
genotyping
transgenic
PCR
reverse-transcription PCR

Software Mentioned

BLAST
TASSER
Clustal W

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