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Related Concept Videos

Conservation of Protein Domains Over Different Proteins02:26

Conservation of Protein Domains Over Different Proteins

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Identification of Footprints of RNA:Protein Complexes via RNA Immunoprecipitation in Tandem Followed by Sequencing (RIPiT-Seq)
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Reiterated WG/GW motifs form functionally and evolutionarily conserved ARGONAUTE-binding platforms in RNAi-related

Mahmoud El-Shami1, Dominique Pontier, Sylvie Lahmy

  • 1Laboratoire Génome et Développement de Plantes, Centre National de la Recherche Scientifique/Institut de Recherche pour le Développement/Université de Perpignan 5096, 66860 Perpignan Cedex, France.

Genes & Development
|October 17, 2007
PubMed
Summary

RNA Polymerase IVb (PolIVb) distinguishes itself from PolIVa in Arabidopsis via its extended C-terminal domain (CTD). This CTD contains WG/GW motifs essential for Argonaute binding and RNA-directed DNA methylation (RdDM).

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PAR-CliP - A Method to Identify Transcriptome-wide the Binding Sites of RNA Binding Proteins

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Area of Science:

  • Molecular Biology
  • Epigenetics
  • Plant Science

Background:

  • Two forms of RNA Polymerase IV (PolIVa/PolIVb) are involved in RNA-directed DNA methylation (RdDM) in Arabidopsis.
  • Current models suggest PolIVb has a unique role due to Argonaute4 (AGO4) association with its largest subunit's C-terminal domain (CTD).

Purpose of the Study:

  • To investigate the functional role of the extended CTD in PolIVb.
  • To determine the mechanism by which PolIVb distinguishes itself from PolIVa in the RdDM pathway.

Main Methods:

  • Site-specific mutagenesis of the AtNRPD1b protein.
  • Domain-swapping experiments between Arabidopsis NRPD1b and human GW182.
  • Analysis of Argonaute-binding capacity and RdDM function.

Main Results:

  • The extended CTD of NRPD1b-type proteins possesses conserved Argonaute-binding capacity.
  • A WG/GW-rich region within the CTD is crucial for distinguishing Pol IVb from Pol IVa.
  • This WG/GW-rich region is essential for the RdDM process.
  • Evolutionarily conserved WG/GW motifs act as Argonaute-binding platforms in RNA interference (RNAi) components.

Conclusions:

  • The extended CTD of PolIVb, featuring WG/GW motifs, is a key determinant of its distinct function in RdDM.
  • These motifs represent conserved Argonaute-binding platforms critical for RNAi pathways across species.