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LMB Seminar: Mechanisms of centriole assembly

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Understanding the organizing principles driving the assembly of cellular organelles and of protein machines is a fundamental pursuit in biology. The centriole is a microtubule-based organelle that is essential for the formation of flagella, cilia and centrosomes. The centriole has a signature nine-fold symmetrical arrangement of microtubules organized around a likewise symmetrical cartwheel. The cartwheel comprises a stack of ring-containing elements that each accommodates 9 homodimers of SAS -6 proteins. Although important advances have been made in recent years, understanding of centriole assembly remains incomplete, partly due to the lack of cell-free assays to probe the dynamics of the process at nanometric scale. To fill this gap, we have developed a high-speed atomic force microscopy (AFM) to reconstitute and monitor the assembly dynamics of SAS -6 proteins. Moreover, to develop tools to dissect and modulate centriole assembly, we have identified monobodies that recognize SAS -6 proteins with high affinity. Our analysis established that some of these monobodies prevent the assembly of ring-containing elements in vitro and of the centriole organelle in cellulo. Together, these approaches provide novel insights into the mechanisms governing the assembly of the fundamental centriole organelle.

This talk is part of the MRC LMB Seminar Series series.

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