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Related Concept Videos

Protein Transport to the Outer Chloroplast Membrane01:11

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Chloroplast outer membrane proteins encoded by the nucleus are synthesized in the cytosol. Soon after synthesis, they bind cytosolic factors such as 14-3-3 protein and the Hsp70 chaperones that keep these precursors in an unfolded state until their translocation.
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Proteins targeted to the inner chloroplast membrane, or plastid proteins, are transported by two general pathways: the stop-transfer and the re-insertion or post-import pathways. Most plastid proteins carry N-terminal transit sequences and internal import sequences targeting it to the specific chloroplast subcompartment. Proteins targeted by the stop-transfer pathway have internal hydrophobic sequences that inhibit their translocation into the stroma. As a result, these precursors are arrested...
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Chloroplasts are triple membrane structures with an outer membrane, an inner membrane, and a thylakoid membrane, each containing distinct metabolite transporters, membrane translocons, and enzymes. Appropriate sorting and translocating these proteins to their correct membrane systems is essential for chloroplast function.
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mRNA Interactome Capture from Plant Protoplasts
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Full Length Transcriptome Highlights the Coordination of Plastid Transcript Processing.

Marine Guilcher1,2, Arnaud Liehrmann1,2,3, Chloé Seyman3

  • 1Institute of Plant Sciences Paris-Saclay (IPS2), Université Paris-Saclay, CNRS, INRAE, Université Evry, 91405 Orsay, France.

International Journal of Molecular Sciences
|October 23, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces nanopore sequencing to analyze the complex plastid transcriptome in Arabidopsis thaliana, revealing new insights into RNA processing and maturation events.

Keywords:
Arabidopsis thalianaco-maturationnanoporeplastidpost-transcriptional

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Area of Science:

  • Plant molecular biology
  • Genomics
  • Transcriptomics

Background:

  • Plastid gene expression is complex, involving multiple post-transcriptional steps and producing diverse RNA isoforms.
  • Short-read RNA sequencing (RNA-Seq) has limitations in capturing full-length plastid transcripts, hindering a complete understanding of gene expression regulation.

Purpose of the Study:

  • To develop and apply a nanopore sequencing protocol for comprehensive analysis of the Arabidopsis thaliana plastid transcriptome.
  • To investigate the interplay of post-transcriptional modifications like editing and splicing in plastid gene expression.

Main Methods:

  • Development of a protocol for nanopore sequencing of plastid RNA from Arabidopsis thaliana leaves.
  • Strand-specific read mapping to the chloroplast genome.
  • Application of virtual Northern blots to analyze transcript complexity.
  • Analysis of co-occurrence and chronology of RNA editing and splicing events.

Main Results:

  • Successfully recapitulated major aspects of plastid transcriptome complexity, including polygenic transcripts and multiple isoforms.
  • Identified 42 pairs of RNA editing and splicing events that are not independent.
  • Observed a preferential order of maturation events, with splicing occurring after most editing sites.

Conclusions:

  • Nanopore sequencing provides a powerful tool for studying the full complexity of the plastid transcriptome.
  • The findings reveal coordinated regulation of post-transcriptional maturation events in plastids.
  • A chronological order of editing and splicing suggests a specific pathway for plastid RNA maturation.