4.8 Article

Assembly of a dsRNA synthesizing complex: RNA-DEPENDENT RNA POLYMERASE 2 contacts the largest subunit of NUCLEAR RNA POLYMERASE IV

Publisher

NATL ACAD SCIENCES
DOI: 10.1073/pnas.2019276118

Keywords

RNA-directed DNA methylation; gene silencing; Pol IV; RDR2; RNA interference

Funding

  1. NIH [GM077590]
  2. Howard Hughes Medical Institute

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In plants, transcription of selfish genetic elements is suppressed by RNA-directed DNA methylation. The process is guided by siRNAs synthesized by Pol IV and RDR2, with the catalytic regions of the two enzymes suggesting a close coupling in their activities.
In plants, transcription of selfish genetic elements such as transposons and DNA viruses is suppressed by RNA-directed DNA methylation. This process is guided by 24-nt short-interfering RNAs (siRNAs) whose double-stranded precursors are synthesized by DNA-dependent NUCLEAR RNA POLYMERASE IV (Pol IV) and RNA-DEPENDENT RNA POLYMERASE 2 (RDR2). Pol IV and RDR2 coimmunoprecipitate, and their activities are tightly coupled, yet the basis for their association is unknown. Here, we show that an interval near the RDR2 active site contacts the Pol IV catalytic subunit, NRPD1, the largest of Pol IV's 12 subunits. Contacts between the catalytic regions of the two enzymes suggests that RDR2 is positioned to rapidly engage the free 3' ends of Pol IV transcripts and convert these single-stranded transcripts into double-stranded RNAs (dsRNAs).

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