title: Subdiffraction-resolution fluorescence microscopy reveals a domain of the centrosome critical for pericentriolar material organization.
abstract: As the main microtubule-organizing centre in animal cells, the centrosome has a fundamental role in cell function. Surrounding the centrioles, the pericentriolar material (PCM) provides a dynamic platform for nucleating microtubules. Although the importance of the PCM is established, its amorphous electron-dense nature has made it refractory to structural investigation. By using SIM and STORM subdiffraction-resolution microscopies to visualize proteins critical for centrosome maturation, we demonstrate that the PCM is organized into two main structural domains: a layer juxtaposed to the centriole wall, and proteins extending farther away from the centriole organized in a matrix. Analysis of Pericentrin-like protein (PLP) reveals that its carboxy terminus is positioned at the centriole wall, it radiates outwards into the matrix and is organized in clusters having quasi-nine-fold symmetry. By RNA-mediated interference (RNAi), we show that PLP fibrils are required for interphase recruitment and proper mitotic assembly of the PCM matrix.
authors.affiliation: Department of Biochemistry and Biophysics and the Howard Hughes Medical Institute, University of California San Francisco, San Francisco, California 94158, USA.
authors.forename: V
authors.initials: V
authors.lastname: Mennella
authors.forename: B
authors.initials: B
authors.lastname: Keszthelyi
authors.forename: K L
authors.initials: KL
authors.lastname: McDonald
authors.forename: B
authors.initials: B
authors.lastname: Chhun
authors.forename: F
authors.initials: F
authors.lastname: Kan
authors.forename: G C
authors.initials: GC
authors.lastname: Rogers
authors.forename: B
authors.initials: B
authors.lastname: Huang
authors.forename: D A
authors.initials: DA
authors.lastname: Agard
mesh_terms: D015153:Blotting, Western; D018797:Cell Cycle Proteins; D002460:Cell Line; D002502:Centrioles / Q000378:metabolism*; D018385:Centrosome / Q000378:metabolism*; D006367:HeLa Cells; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D008853:Microscopy; D008856:Microscopy, Fluorescence / Q000379:methods*; D008870:Microtubules / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000378:metabolism; D034622:RNA Interference
chemical_list: C405929:CDK5RAP2 protein, human; D018797:Cell Cycle Proteins; D047908:Intracellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins