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SID-1 is a dsRNA-selective dsRNA-gated channel.

Joseph D Shih1, Craig P Hunter

  • 1Department of Molecular and Cellular Biology, Harvard University, Cambridge, Massachusetts 02138, USA.

RNA (New York, N.Y.)
|April 9, 2011
PubMed
Summary

The transmembrane protein SID-1 facilitates systemic RNA interference (RNAi) by transporting double-stranded RNA (dsRNA) between cells. This study reveals SID-1 acts as a dsRNA-gated channel, crucial for RNAi signaling and potentially other RNA-based biological processes.

Area of Science:

  • Molecular Biology
  • Genetics
  • Cell Biology

Background:

  • Systemic RNA interference (RNAi) in Caenorhabditis elegans relies on the transmembrane protein SID-1 for intercellular transport of silencing signals.
  • SID-1 expression in Drosophila S2 cells facilitates passive uptake of double-stranded RNA (dsRNA), suggesting a channel function for dsRNA transport.

Purpose of the Study:

  • To elucidate the mechanism of nucleic acid transport by SID-1.
  • To determine if SID-1 functions as a dsRNA-gated channel.
  • To investigate the role of the RNA-induced silencing complex (RISC) in dsRNA retention and explore the transport of natural RNA mimics by SID-1.

Main Methods:

  • Expressing Caenorhabditis elegans SID-1 in Drosophila S2 cells.
  • Measuring membrane conductance changes upon dsRNA addition.

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  • Assessing dsRNA retention by the RNA-induced silencing complex (RISC).
  • Testing the transport of hairpin RNA and pre-microRNA by SID-1.

    • Main Results:

    • SID-1-mediated nucleic acid transport is specific for dsRNA.
    • dsRNA binding to SID-1 alters membrane conductance, indicating a dsRNA-gated channel mechanism.
    • RISC is necessary to prevent dsRNA export, without apparent processing into siRNAs.
    • SID-1 transports natural RNA molecules like hairpin RNA and pre-microRNA.

    Conclusions:

    • SID-1 functions as a dsRNA-gated channel protein, mediating bidirectional dsRNA transport.
    • RISC plays a role in retaining imported dsRNA, essential for RNAi signaling.
    • SID-1 can transport various dsRNA-containing molecules, offering insights into endogenous animal RNA signaling.