C1D and hMtr4p associate with the human exosome subunit PM/Scl-100 and are involved in pre-rRNA processing.
The exosome is a complex of 3'-5' exoribonucleases and RNA-binding proteins, which is involved in processing or degradation of different classes of RNA. Previously, the characterization of purified exosome complexes from yeast and human cells suggested that C1D and KIAA0052/hMtr4p are associated with the exosome and thus might regulate its functional activities. Subcellular localization experiments demonstrated that C1D and KIAA0052/hMtr4p co-localize with exosome subunit PM/Scl-100 in the nucleoli of HEp-2 cells. Additionally, the nucleolar accumulation of C1D appeared to be dependent on PM/Scl-100. Protein-protein interaction studies showed that C1D binds to PM/Scl-100, whereas KIAA0052/hMtr4p was found to interact with MPP6, a previously identified exosome-associated protein. Moreover, we demonstrate that C1D, MPP6 and PM/Scl-100 form a stable trimeric complex in vitro. Knock-down of C1D, MPP6 and KIAA0052/hMtr4p by RNAi resulted in the accumulation of 3'-extended 5.8S rRNA precursors, showing that these proteins are required for rRNA processing. Interestingly, C1D appeared to contain RNA-binding activity with a potential preference for structured RNAs. Taken together, our results are consistent with a role for the exosome-associated proteins C1D, MPP6 and KIAA052/hMtr4p in the recruitment of the exosome to pre-rRNA to mediate the 3' end processing of the 5.8S rRNA.
Schilders
Geurt
G
Department of Biomolecular Chemistry, Nijmegen Center for Molecular Life Sciences, Institute for Molecules and Materials, Radboud University Nijmegen, Nijmegen, The Netherlands.
van Dijk
Erwin
E
Pruijn
Ger J M
GJ
D002460:Cell Line; D002466:Cell Nucleolus / Q000737:chemistry / Q000201:enzymology; D056970:Co-Repressor Proteins; D005095:Exoribonucleases / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D063326:Exosome Multienzyme Ribonuclease Complex; D006801:Humans; D009687:Nuclear Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D026721:RNA 3' End Processing; D020365:RNA Helicases / Q000032:analysis / Q000378:metabolism / Q000502:physiology*; D012322:RNA Precursors / Q000378:metabolism*; D012340:RNA, Ribosomal, 5.8S / Q000378:metabolism*; D016601:RNA-Binding Proteins / Q000378:metabolism; D012097:Repressor Proteins / Q000032:analysis / Q000378:metabolism / Q000502:physiology*
C117400:C1D protein, human; D056970:Co-Repressor Proteins; C507180:M-phase phosphoprotein 6, human; D009687:Nuclear Proteins; D012322:RNA Precursors; D012340:RNA, Ribosomal, 5.8S; D016601:RNA-Binding Proteins; D012097:Repressor Proteins; D005095:Exoribonucleases; D063326:Exosome Multienzyme Ribonuclease Complex; C077044:EXOSC10 protein, human; C512513:MTREX protein, human; D020365:RNA Helicases