Human cell growth requires a functional cytoplasmic exosome, which is involved in various mRNA decay pathways

The human exosome is a 3'-5' exoribonuclease complex that functions both in the nucleus and in the cytoplasm to either degrade or process RNA. Little is known yet about potential differences among core exosome complexes in these different cellular compartments and the roles of the individual subunits in maintaining a stable and functional complex. Glycerol gradient sedimentation analyses indicated that a significant subset of nuclear exosomes is present in much larger complexes (60-80S) than the cytoplasmic exosomes (similar to 10S). Interestingly, siRNA-mediated knock- down experiments indicated that the cytoplasmic exosome is down- regulated much more efficiently than the nuclear exosome. In addition, we observed that knockdown of hRrp41p or hRrp4p but not PM/ Scl-100 or PM/ Scl-75 leads to codepletion of other subunits. Nevertheless, PM/ Scl-100 and PM/ Scl- 75 are required to maintain normal levels of three different mRNA reporters: a wild- type beta-globin mRNA, a b- globin mRNA containing an AU- rich ( ARE) instability element, and a b- globin mRNA bearing a premature termination codon ( PTC). The increased levels of ARE- and the PTC-containing mRNAs upon down- regulation of the different exosome subunits, in particular PM/ Scl- 100, appeared to be due to decreased turnover rates. These results indicate that, although not required for exosome stability, PM/ Scl-100 and PM/ Scl-75 are involved in mRNA degradation, either as essential subunits of a functional exosome complex or as exosome- independent proteins.