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  • Direct interaction of Plk4 with STIL ensures formation of a single procentriole per parental centriole.

Direct interaction of Plk4 with STIL ensures formation of a single procentriole per parental centriole.

Nature communications (2014-10-25)
Midori Ohta, Tomoko Ashikawa, Yuka Nozaki, Hiroko Kozuka-Hata, Hidemasa Goto, Masaki Inagaki, Masaaki Oyama, Daiju Kitagawa
ABSTRACT

Formation of one procentriole next to each pre-existing centriole is essential for centrosome duplication, robust bipolar spindle assembly and maintenance of genome integrity. However, the mechanisms maintaining strict control over centriole copy number are incompletely understood. Here we show that Plk4 and STIL, the key regulators of centriole formation, form a protein complex that provides a scaffold for recruiting HsSAS-6, a major component of the centriolar cartwheel, at the onset of procentriole formation. Furthermore, we demonstrate that phosphorylation of STIL by Plk4 facilitates the STIL/HsSAS-6 interaction and centriolar loading of HsSAS-6. We also provide evidence that negative feedback by centriolar STIL regulates bimodal centriolar distribution of Plk4 and seemingly restricts occurrence of procentriole formation to one site on each parental centriole. Overall, these findings suggest a mechanism whereby coordinated action of three critical factors ensures formation of a single procentriole per parental centriole.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Centrin Antibody, clone 20H5, clone 20H5, from mouse
Sigma-Aldrich
Monoclonal ANTI-FLAG® M2 antibody produced in mouse, 1 mg/mL, clone M2, affinity isolated antibody, buffered aqueous solution (50% glycerol, 10 mM sodium phosphate, and 150 mM NaCl, pH 7.4)
Sigma-Aldrich
Monoclonal Anti-α-Tubulin antibody produced in mouse, clone DM1A, ascites fluid
Millipore
ANTI-FLAG® M2 Affinity Gel, purified immunoglobulin, buffered aqueous glycerol solution