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Identification of Alternative Splicing and Polyadenylation in RNA-seq Data
Published on: June 24, 2021
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Modulation of Auxin Signaling and Development by Polyadenylation Machinery
Wei Zeng1,2, Xinhua Dai3, Jing Sun4
1Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, Beijing 100093, China.
Plant Physiology
|November 30, 2018
Summary
Cleavage stimulation Factor 77 (AtCstF77) in Arabidopsis controls polyadenylation site selection, impacting auxin signaling and plant development. Disruption of AtCstF77 causes developmental defects and altered auxin responses by shifting mRNA polyadenylation sites.
Area of Science:
- Plant Molecular Biology
- Gene Expression Regulation
- Arabidopsis thaliana Research
Background:
- Polyadenylation is crucial for mRNA processing, influencing stability, transport, and translation.
- The Cleavage stimulation Factor 77 (AtCstF77) is a key component of the pre-mRNA 3'-end polyadenylation machinery in plants.
- Previous understanding of AtCstF77's role in Arabidopsis development and gene regulation was incomplete.
Purpose of the Study:
- To investigate the function of AtCstF77 in Arabidopsis, particularly its role in polyadenylation site (PAS) selection.
- To determine the impact of AtCstF77 on auxin signaling pathways and plant development.
- To elucidate the molecular mechanisms by which AtCstF77 influences gene expression.
Main Methods:
- Analysis of Arabidopsis mutants with disrupted AtCstF77 genes.
- Assessment of auxin sensitivity and reporter gene expression (DR5-GFP).
- RNA sequencing to identify polyadenylation site shifts across the transcriptome.
- Genetic interaction studies with auxin receptor mutants.
- Expression analysis of auxin signaling and homeostasis genes.
Main Results:
- Disruption of AtCstF77 led to reduced auxin sensitivity and decreased DR5-GFP expression.
- Null mutations in AtCstF77 caused severe developmental defects, though not lethal.
- Polyadenylation site (PAS) shifts were observed in approximately 2,400 genes in the cstf77-2 mutant, predominantly from proximal to distal sites.
- Several auxin signaling genes exhibited altered PASs in the cstf77-2 mutant, with increased expression of AUXIN RESISTANT 2/INDOLE-3-ACETIC ACID 7 contributing to auxin resistance.
- Genetic interactions with auxin receptor mutants confirmed the link between polyadenylation and auxin signaling.
Conclusions:
- AtCstF77 plays critical and pleiotropic roles in Arabidopsis development.
- AtCstF77 regulates auxin responses, primarily by controlling PAS selection in transcripts of auxin signaling components.
- The findings highlight the importance of polyadenylation in integrating gene expression regulation with plant hormone signaling.
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