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Published on: December 20, 2014
RBM10 regulates centriole duplication in HepG2 cells by ectopically assembling PLK4-STIL complexes in the nucleus
Hiroyuki Kunimoto1, Akira Inoue1, Hirotada Kojima1
1Department of Immunology, Graduate School of Medicine, Osaka City University, Osaka, Japan.
Abstract:
RNA-binding motif protein 10 (RBM10) primarily regulates alternative splicing of certain genes. Loss-of-function mutations in RBM10 have been frequently reported in patients with various cancers. However, how RBM10 levels affect cell proliferation and tumorigenesis remains unknown. To elucidate the role of RBM10 in cell proliferation, we established HepG2-RBM10 knockout cell lines and derivative doxycycline-inducible RBM10-expressing cells. RBM10 over-expression caused growth arrest in the M phase with a monopolar spindle because of impaired centriole duplication. Two RBM10 splicing mutants, one with F345A/F347A and the other with only the C-terminal half (401-930), were sufficient to cause growth arrest, whereas an RBM10 mutant with cytoplasmic localization forced by an NES did not show growth arrest. RBM10 over-expression induced the formation of many large nuclear domains containing RBM10, PLK4, STIL and SAS6, which are the regulatory proteins involved in centriole duplication. Consistently, the centrioles in the RBM10-over-expressing HepG2 cells lost PLK4 and STIL, accounting for the unsuccessful centriole duplication. In contrast, RBM10 depletion resulted in elevated levels of cytoplasmic PLK4 with a concomitant increase in the number of centrioles in HepG2 cells but not in A549 cells. Thus, nuclear RBM10 regulates normal chromosomal division in a cell-type-specific manner, independent of alternative RNA splicing.
Insights
RNA-binding motif protein 10 (RBM10) over-expression halts cell division by impairing centriole duplication, independent of its splicing role. Nuclear RBM10 controls chromosome segregation in a cell-type-specific manner.
Area of Science:
- Cell Biology
- Molecular Biology
- Cancer Research
Background:
- RNA-binding motif protein 10 (RBM10) mutations are linked to various cancers.
- The precise role of RBM10 in cell proliferation and tumorigenesis is not fully understood.
Purpose of the Study:
- To investigate the function of RBM10 in cell proliferation and its impact on cell division.
- To elucidate the mechanism by which RBM10 influences centriole duplication and chromosomal segregation.
Main Methods:
- Establishment of HepG2 cell lines with RBM10 knockout and inducible RBM10 expression.
- Analysis of cell cycle progression, spindle formation, and centriole duplication upon RBM10 manipulation.
- Investigation of RBM10 localization and interaction with centriole duplication proteins (PLK4, STIL, SAS6).
Main Results:
- RBM10 over-expression leads to M phase growth arrest due to impaired centriole duplication and monopolar spindles.
- Nuclear RBM10, but not cytoplasmic RBM10, induced growth arrest.
- RBM10 over-expression formed nuclear domains with centriole duplication proteins, disrupting centriole PLK4 and STIL.
- RBM10 depletion increased cytoplasmic PLK4 and centriole numbers in HepG2 cells, but not A549 cells.
Conclusions:
- Nuclear RBM10 regulates centriole duplication and chromosomal segregation in a cell-type-specific manner.
- RBM10's role in cell division is independent of its alternative RNA splicing function.
- RBM10's impact on cell proliferation suggests a potential role in cancer development.
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