Format

Send to

Choose Destination
See comment in PubMed Commons below
Sci Rep. 2016 Apr 14;6:24296. doi: 10.1038/srep24296.

Molecular basis of the STIL coiled coil oligomerization explains its requirement for de-novo formation of centrosomes in mammalian cells.

Author information

1
Sheba Cancer Research Center and the Edmond and Lily Safra Children Hospital, Sheba Medical Center, Tel-Hashomer 52621, Israel.
2
Department of molecular genetics and biochemistry, Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel.
3
Institute of Chemistry, the Hebrew University of Jerusalem, Safra Campus, Givat Ram, Jerusalem 91904, Israel.

Abstract

The STIL protein is essential for centriole replication and for the non-templated, de novo centriole biogenesis that is required for mammalian embryogenesis. Here we performed quantitative biophysical and structural analysis of the central short coiled coil domain (CCD) of STIL that is critical for its function. Using biophysical, biochemical and cell biology approaches, we identified the specific residues in the CCD that mediate the oligomerization, centrosomal localization and protein interactions of STIL. We characterized the structural properties of the coiled coil peptide using circular dichroism spectroscopy and size exclusion chromatography. We identified two regions in this domain, containing eight hydrophobic residues, which mediate the coiled coil oligomerization. Mutations in these residues destabilized the coiled coil thermodynamically but in most cases did not affect its secondary structure. Reconstituting mouse embryonic fibroblasts lacking endogenous Stil, we show that STIL oligomerization mediated by these residues is not only important for the centrosomal functions of STIL during the canonical duplication process but also for de-novo formation of centrosomes.

PMID:
27075531
PMCID:
PMC4830966
DOI:
10.1038/srep24296
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Nature Publishing Group Icon for PubMed Central
    Loading ...
    Support Center