Dominant-negative Pes1 mutants inhibit ribosomal RNA processing and cell proliferation via incorporation into the PeBoW-complex.
The nucleolar PeBoW-complex, consisting of Pes1, Bop1 and WDR12, is essential for cell proliferation and processing of ribosomal RNA in mammalian cells. Here we have analysed the physical and functional interactions of Pes1 deletion mutants with the PeBoW-complex. Pes1 mutants M1 and M5, with N- and C-terminal truncations, respectively, displayed a dominant-negative phenotype. Both mutants showed nucleolar localization, blocked processing of the 36S/32S precursors to mature 28S rRNA, inhibited cell proliferation, and induced high p53 levels in proliferating, but not in resting cells. Mutant M1 and M5 proteins associated with large pre-ribosomal complexes and co-immunoprecipitated Bop1 and WDR12 proteins indicating their proper incorporation into the PeBoW-complex. We conclude that the dominant-negative effect of the M1 and M5 mutants is mediated by the impaired function of the PeBoW-complex.
Grimm
Thomas
T
Institute of Clinical Molecular Biology and Tumour Genetics, GSF Research Centre for Environment and Health, Marchioninistrasse 25, 81377 Munich, Germany.
Hölzel
Michael
M
Rohrmoser
Michaela
M
Harasim
Thomas
T
Malamoussi
Anastassia
A
Gruber-Eber
Anita
A
Kremmer
Elisabeth
E
Eick
Dirk
D
D000818:Animals; D002453:Cell Cycle; D002460:Cell Line; D002466:Cell Nucleolus / Q000737:chemistry; D049109:Cell Proliferation; D003001:Cloning, Molecular; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D011506:Proteins / Q000032:analysis / Q000235:genetics / Q000378:metabolism*; D012322:RNA Precursors / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D012335:RNA, Ribosomal / Q000378:metabolism*; D016601:RNA-Binding Proteins; D051381:Rats; D012270:Ribosomes / Q000378:metabolism; D017384:Sequence Deletion; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D009687:Nuclear Proteins; C484463:PES1 protein, human; D011506:Proteins; D012322:RNA Precursors; D012335:RNA, Ribosomal; D016601:RNA-Binding Proteins; D016159:Tumor Suppressor Protein p53