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Originally published as MBC in Press, 10.1091/mbc.E07-01-0070 on June 27, 2007

Vol. 18, Issue 9, 3375-3387, September 2007

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Small Interfering RNA-mediated Silencing Induces Target-dependent Assembly of GW/P BodiesFormula

Shangli Lian*, Marvin J. Fritzler{dagger}, Joseph Katz{ddagger}, Takashi Hamazaki§, Naohiro Terada§, Minoru Satoh§,||, and Edward K.L. Chan*

Departments of *Oral Biology, {ddagger}Oral Medicine, and §Pathology, Immunology, and Laboratory Medicine, and ||Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Florida, Gainesville, FL 32610; and {dagger}Department of Biochemistry and Molecular Biology, University of Calgary, Alberta, Canada T2N 1N4

Submitted January 26, 2007; Revised June 18, 2007; Accepted June 19, 2007
Monitoring Editor: A. Gregory Matera

Gene silencing using small interfering RNA (siRNA) is a valuable laboratory tool and a promising approach to therapeutics for a variety of human diseases. Recently, RNA interference (RNAi) has been linked to cytoplasmic GW bodies (GWB). However, the correlation between RNAi and the formation of GWB, also known as mammalian processing bodies, remains unclear. In this report, we show that transfection of functional siRNA induced larger and greater numbers of GWB. This siRNA-induced increase of GWB depended on the endogenous expression of the target mRNA. Knockdown of GW182 or Ago2 demonstrated that the siRNA-induced increase of GWB required these two proteins and correlated with RNAi. Furthermore, knockdown of rck/p54 or LSm1 did not prevent the reassembly of GWB that were induced by and correlated with siRNA-mediated RNA silencing. We propose that RNAi is a key regulatory mechanism for the assembly of GWB, and in some cases, GWB may serve as markers for RNAi in mammalian cells.

This was published online ahead of print in MBC in Press (http://www.molbiolcell.org/cgi/doi/10.1091/mbc.E07-01-0070) on June 27, 2007.

Formula The online version of this article contains supplemental material at MBC Online (http://www.molbiolcell.org).

Address correspondence to: Edward K.L. Chan (echan{at}ufl.edu).

Abbreviations used: GWB, GW bodies; IIF, indirect immunofluorescence; miRNA, microRNA; P bodies, processing bodies; RISC, RNA-induced silencing complex; SG, stress granules.




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