Hydrolysis rates of different small interfering RNAs (siRNAs) by the RNA silencing promoter complex, C3PO, determines their regulation by phospholipase Cβ.

The Journal of Biological Chemistry
Shriya SahuSuzanne Scarlata

Abstract

C3PO plays a key role in promoting RNA-induced gene silencing. C3PO consists of two subunits of the endonuclease translin-associated factor X (TRAX) and six subunits of the nucleotide-binding protein translin. We have found that TRAX binds strongly to phospholipase Cβ (PLCβ), which transmits G protein signals from many hormones and sensory inputs. The association between PLCβ and TRAX is thought to underlie the ability of PLCβ to reverse gene silencing by small interfering RNAs. However, this reversal only occurs for some genes (e.g. GAPDH and LDH) but not others (e.g. Hsp90 and cyclophilin A). To understand this specificity, we carried out studies using fluorescence-based methods. In cells, we find that PLCβ, TRAX, and their complexes are identically distributed through the cytosol suggesting that selectivity is not due to large scale sequestration of either the free or complexed proteins. Using purified proteins, we find that PLCβ binds ∼5-fold more weakly to translin than to TRAX but ∼2-fold more strongly to C3PO. PLCβ does not alter TRAX-translin assembly to C3PO, and brightness studies suggest one PLCβ binds to one C3PO octamer without a change in the number of TRAX/translin molecules suggesting that PLCβ binds to an exter...Continue Reading

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Citations

Jan 10, 2016·Advances in Biological Regulation·Suzanne ScarlataUrszula Golebiewska
Jul 18, 2017·Protein Science : a Publication of the Protein Society·Shriya SahuSuzanne Scarlata
Dec 24, 2019·Protein Science : a Publication of the Protein Society·Lela JacksonSuzanne Scarlata
Feb 11, 2016·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Finly PhilipSuzanne Scarlata
Oct 30, 2016·The Journal of Biological Chemistry·Osama Garwain, Suzanne Scarlata

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