Formation of a new centriole adjacent to a pre-existing centriole occurs only once per cell cycle. Despite being crucial for genome integrity, the mechanisms controlling centriole biogenesis remain elusive. Here, we identify RBM14 as a novel suppressor of assembly of centriolar protein complexes. Depletion of RBM14 in human cells induces ectopic formation of centriolar protein complexes through function of the STIL/CPAP complex. Intriguingly, the formation of such structures seems not to require the cartwheel structure that normally acts as a scaffold for centriole formation, whereas they can retain pericentriolar material and microtubule nucleation activity. Moreover, we find that, upon RBM14 depletion, a part of the ectopic centriolar protein complexes in turn assemble into structures more akin to centrioles, presumably by incorporating HsSAS-6, a cartwheel component, and cause multipolar spindle formation. We further demonstrate that such structures assemble in the cytoplasm even in the presence of pre-existing centrioles. This study sheds light on the possibility that ectopic formation of aberrant structures related to centrioles may contribute to genome instability and tumorigenesis.
Centriole duplication occurs once per cell cycle through the assembly of daughter centrioles on the side wall of pre-existing centrioles. Little is known about the molecules involved in the assembly of new centrioles. Here, we identify CRC70 as a Chlamydomonas protein with an important role in the accumulation of centriole proteins at the site of assembly. CRC70 contains a highly conserved ~50-amino-acid sequence shared by mammalian Cep70 and preferentially localizes to immature centrioles (the procentrioles). This localization is maintained in the mutant bld10, in which centriole formation is blocked before the assembly of centriolar microtubules. RNA interference (RNAi)-mediated knockdown of CRC70 produces flagella-less cells and inhibits the recruitment of other centriole components, such as SAS-6 and Bld10p to the centriole. Overexpression of CRC70 induces an accumulation of these proteins in discrete spots in the cytoplasm. Overexpression of EGFP-tagged CRC70 in mouse NIH3T3 cells causes the formation of structures apparently related to centrioles. These findings suggest that CRC70 is a member of a conserved protein family and functions as a scaffold for the assembly of the centriole precursor.
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