Format

Send to

Choose Destination
J Cell Biol. 2014 Mar 3;204(5):697-712. doi: 10.1083/jcb.201307049.

Mechanisms of HsSAS-6 assembly promoting centriole formation in human cells.

Author information

1
Swiss Institute for Experimental Cancer Research, School of Life Sciences, 2 Laboratory for Experimental Biophysics, and 3 Institute for Science and Chemical Engineering, Swiss Federal Institute of Technology, 1015 Lausanne, Switzerland.

Abstract

SAS-6 proteins are thought to impart the ninefold symmetry of centrioles, but the mechanisms by which their assembly occurs within cells remain elusive. In this paper, we provide evidence that the N-terminal, coiled-coil, and C-terminal domains of HsSAS-6 are each required for procentriole formation in human cells. Moreover, the coiled coil is necessary and sufficient to mediate HsSAS-6 centrosomal targeting. High-resolution imaging reveals that GFP-tagged HsSAS-6 variants localize in a torus around the base of the parental centriole before S phase, perhaps indicative of an initial loading platform. Moreover, fluorescence recovery after photobleaching analysis demonstrates that HsSAS-6 is immobilized progressively at centrosomes during cell cycle progression. Using fluorescence correlation spectroscopy and three-dimensional stochastic optical reconstruction microscopy, we uncover that HsSAS-6 is present in the cytoplasm primarily as a homodimer and that its oligomerization into a ninefold symmetrical ring occurs at centrioles. Together, our findings lead us to propose a mechanism whereby HsSAS-6 homodimers are targeted to centrosomes where the local environment and high concentration of HsSAS-6 promote oligomerization, thus initiating procentriole formation.

PMID:
24590172
PMCID:
PMC3941056
DOI:
10.1083/jcb.201307049
[Indexed for MEDLINE]
Free PMC Article

Supplemental Content

Full text links

Icon for HighWire Icon for PubMed Central
Loading ...
Support Center