TTP and BRF proteins nucleate processing body formation to silence mRNAs with AU-rich elements

In mammalian cells, mRNAs with AU-rich elements (AREs) are targeted for translational silencing and rapid degradation. Here we present evidence that in human cells the proteins Tristetraprolin (TTP) and BRF-1 deliver ARE-mRNAs to processing bodies (PBs), cytoplasmic assemblies of mRNAs, and associated factors that promote translational silencing and mRNA decay. First, depletion of endogenous TTP and BRF proteins, or overexpression of dominant-negative mutant TTP proteins, impairs the localization of reporter ARE-mRNAs in PBs. Second, TTP and BRF-1 localize tethered mRNAs to PBs. Third, TTP can nucleate PB formation on untranslated mRNAs even when other mRNAs are trapped in polysomes by cycloheximide treatment. ARE-mRNA localization in PBs is mediated by the TTP N- and C-terminal domains and occurs downstream from mRNA polysome release, which in itself is not sufficient for mRNA PB localization. The accumulation of ARE-mRNAs in PBs is strongly enhanced when the mRNA decay machinery is rendered limiting by mRNA decay enzyme depletion or TTP/BRF-1 overexpression. Based on these observations, we propose that the PB functions as a reservoir that sequesters ARE-mRNAs from polysomes, thereby silencing ARE-mRNA function even when mRNA decay is delayed. This function of the PB can likely be extended to other mRNA silencing pathways, such as those mediated by microRNAs, premature termination codons, and mRNA deadenylation.