3D-structured illumination microscopy provides novel insight into architecture of human centrosomes
JournalArticle (Originalarbeit in einer wissenschaftlichen Zeitschrift)
 
ID 1280002
Author(s) Sonnen, Katharina F; Schermelleh, Lothar; Leonhardt, Heinrich; Nigg, Erich A
Author(s) at UniBasel Nigg, Erich
Year 2012
Title 3D-structured illumination microscopy provides novel insight into architecture of human centrosomes
Journal Biology open
Volume 1
Number 10
Pages / Article-Number 965-76
Keywords 3D-SIM, Plk4, Centriole, Centrosome, Super-resolution microscopy
Abstract Centrioles are essential for the formation of cilia and flagella. They also form the core of the centrosome, which organizes microtubule arrays important for cell shape, polarity, motility and division. Here, we have used super-resolution 3D-structured illumination microscopy to analyse the spatial relationship of 18 centriole and pericentriolar matrix (PCM) components of human centrosomes at different cell cycle stages. During mitosis, PCM proteins formed extended networks with interspersed ?-Tubulin. During interphase, most proteins were arranged at specific distances from the walls of centrioles, resulting in ring staining, often with discernible density masses. Through use of site-specific antibodies, we found the C-terminus of Cep152 to be closer to centrioles than the N-terminus, illustrating the power of 3D-SIM to study protein disposition. Appendage proteins showed rings with multiple density masses, and the number of these masses was strongly reduced during mitosis. At the proximal end of centrioles, Sas-6 formed a dot at the site of daughter centriole assembly, consistent with its role in cartwheel formation. Plk4 and STIL co-localized with Sas-6, but Cep135 was associated mostly with mother centrioles. Remarkably, Plk4 formed a dot on the surface of the mother centriole before Sas-6 staining became detectable, indicating that Plk4 constitutes an early marker for the site of nascent centriole formation. Our study provides novel insights into the architecture of human centrosomes and illustrates the power of super-resolution microscopy in revealing the relative localization of centriole and PCM proteins in unprecedented detail.
Publisher The Company of Biologists
ISSN/ISBN 2046-6390
URL http://dx.doi.org/10.1242/bio.20122337
edoc-URL http://edoc.unibas.ch/dok/A6008535
Full Text on edoc No
Digital Object Identifier DOI 10.1242/bio.20122337
PubMed ID http://www.ncbi.nlm.nih.gov/pubmed/23213374
ISI-Number WOS:000209205500005
Document type (ISI) Journal Article
 
   

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