%0 Journal Article %1 Wueseke2016 %A Wueseke, Oliver %A Zwicker, David %A Schwager, Anne %A Wong, Yao Liang %A Oegema, Karen %A Jülicher, Frank %A Hyman, Anthony A %A Woodruff, Jeffrey B %D 2016 %J Biol. Open %K Centrosome;_phase_sep._/_drops %P 1431-1440 %R 10.1242/bio.020990 %T Polo kinase phosphorylation determines C. elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation %V 5 %X Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In C. elegans embryos, the mitotic PCM expands by Polo-kinase (PLK-1) phosphorylation-accelerated assembly of SPD-5 molecules into supramolecular scaffolds. However, how PLK-1 phosphorylation regulates SPD-5 assembly is not known. We found that a mutant version of SPD-5 that is insensitive to PLK-1 phosphorylation (SPD-5(4A)) could localize to PCM but was unable to rescue the reduction in PCM size and density when wild-type SPD-5 levels were decreased. In vitro, purified SPD-5(4A) self-assembled into functional supramolecular scaffolds over long time scales, suggesting that phosphorylation only controls the rate of SPD-5 scaffold assembly. Furthermore, the SPD-5 scaffold, once assembled, remained intact and supported microtubule nucleation in the absence of PLK-1 activity in vivo We conclude that Polo Kinase is required for rapid assembly of the PCM scaffold but not for scaffold maintenance or function. Based on this idea, we developed a theoretical model that adequately predicted PCM growth rates in different mutant conditions in vivo We propose that PLK-1 phosphorylation-dependent conversion of SPD-5 into an assembly-competent form underlies PCM formation in vivo and that the rate of this conversion determines final PCM size and density.

@article{Wueseke2016, abstract = {Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In C. elegans embryos, the mitotic PCM expands by Polo-kinase (PLK-1) phosphorylation-accelerated assembly of SPD-5 molecules into supramolecular scaffolds. However, how PLK-1 phosphorylation regulates SPD-5 assembly is not known. We found that a mutant version of SPD-5 that is insensitive to PLK-1 phosphorylation (SPD-5(4A)) could localize to PCM but was unable to rescue the reduction in PCM size and density when wild-type SPD-5 levels were decreased. In vitro, purified SPD-5(4A) self-assembled into functional supramolecular scaffolds over long time scales, suggesting that phosphorylation only controls the rate of SPD-5 scaffold assembly. Furthermore, the SPD-5 scaffold, once assembled, remained intact and supported microtubule nucleation in the absence of PLK-1 activity in vivo We conclude that Polo Kinase is required for rapid assembly of the PCM scaffold but not for scaffold maintenance or function. Based on this idea, we developed a theoretical model that adequately predicted PCM growth rates in different mutant conditions in vivo We propose that PLK-1 phosphorylation-dependent conversion of SPD-5 into an assembly-competent form underlies PCM formation in vivo and that the rate of this conversion determines final PCM size and density.}, added-at = {2017-03-02T23:22:01.000+0100}, author = {Wueseke, Oliver and Zwicker, David and Schwager, Anne and Wong, Yao Liang and Oegema, Karen and J{\"u}licher, Frank and Hyman, Anthony A and Woodruff, Jeffrey B}, bdsk-file-1 = {YnBsaXN0MDDUAQIDBAUGJCVYJHZlcnNpb25YJG9iamVjdHNZJGFyY2hpdmVyVCR0b3ASAAGGoKgHCBMUFRYaIVUkbnVsbNMJCgsMDxJXTlMua2V5c1pOUy5vYmplY3RzViRjbGFzc6INDoACgAOiEBGABIAFgAdccmVsYXRpdmVQYXRoWWFsaWFzRGF0YV8QmC4uL0RvY3VtZW50cy9CaWJsaW9ncmFwaHkvUGFwZXJzL1d1ZXNla2UyMDE2IC0gUG9sbyBraW5hc2UgcGhvc3Bob3J5bGF0aW9uIGRldGVybWluZXMgQy4gZWxlZ2FucyBjZW50cm9zb21lIHNpemUgYW5kIGRlbnNpdHkgYnkgYmlhc2luZyBTUEQtNSB0b3dhcmEucGRm0hcLGBlXTlMuZGF0YU8RAvwAAAAAAvwAAgAADE1hY2ludG9zaCBIRAAAAAAAAAAAAAAAAAAAANRxYVhIKwAAABi9gR9XdWVzZWtlMjAxNiAtIFBvbG8gayMxOEM3OTcucGRmAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAGMeX1CqnggAAAAAAAAAAAAEABAAACSAAAAAAAAAAAAAAAAAAAAAGUGFwZXJzABAACAAA1HGnqAAAABEACAAA1CrfwgAAAAEAFAAYvYEAGLzRAAmpegAJqXkABmIvAAIAXU1hY2ludG9zaCBIRDpVc2VyczoAendpY2tlcjoARG9jdW1lbnRzOgBCaWJsaW9ncmFwaHk6AFBhcGVyczoAV3Vlc2VrZTIwMTYgLSBQb2xvIGsjMThDNzk3LnBkZgAADgDwAHcAVwB1AGUAcwBlAGsAZQAyADAAMQA2ACAALQAgAFAAbwBsAG8AIABrAGkAbgBhAHMAZQAgAHAAaABvAHMAcABoAG8AcgB5AGwAYQB0AGkAbwBuACAAZABlAHQAZQByAG0AaQBuAGUAcwAgAEMALgAgAGUAbABlAGcAYQBuAHMAIABjAGUAbgB0AHIAbwBzAG8AbQBlACAAcwBpAHoAZQAgAGEAbgBkACAAZABlAG4AcwBpAHQAeQAgAGIAeQAgAGIAaQBhAHMAaQBuAGcAIABTAFAARAAtADUAIAB0AG8AdwBhAHIAYQAuAHAAZABmAA8AGgAMAE0AYQBjAGkAbgB0AG8AcwBoACAASABEABIAo1VzZXJzL3p3aWNrZXIvRG9jdW1lbnRzL0JpYmxpb2dyYXBoeS9QYXBlcnMvV3Vlc2VrZTIwMTYgLSBQb2xvIGtpbmFzZSBwaG9zcGhvcnlsYXRpb24gZGV0ZXJtaW5lcyBDLiBlbGVnYW5zIGNlbnRyb3NvbWUgc2l6ZSBhbmQgZGVuc2l0eSBieSBiaWFzaW5nIFNQRC01IHRvd2FyYS5wZGYAABMAAS8AABUAAgAO//8AAIAG0hscHR5aJGNsYXNzbmFtZVgkY2xhc3Nlc11OU011dGFibGVEYXRhox0fIFZOU0RhdGFYTlNPYmplY3TSGxwiI1xOU0RpY3Rpb25hcnmiIiBfEA9OU0tleWVkQXJjaGl2ZXLRJidUcm9vdIABAAgAEQAaACMALQAyADcAQABGAE0AVQBgAGcAagBsAG4AcQBzAHUAdwCEAI4BKQEuATYENgQ4BD0ESARRBF8EYwRqBHMEeASFBIgEmgSdBKIAAAAAAAACAQAAAAAAAAAoAAAAAAAAAAAAAAAAAAAEpA==}, bdsk-url-1 = {http://dx.doi.org/10.1242/bio.020990}, biburl = {https://www.bibsonomy.org/bibtex/28c89f305a9ebd3db10b88936194c9da3/david-zwicker}, date-added = {2016-09-07 13:37:53 +0000}, date-modified = {2016-10-18 15:47:49 +0000}, doi = {10.1242/bio.020990}, interhash = {e177bb4f5fdb992c4ded557a6d92f007}, intrahash = {8c89f305a9ebd3db10b88936194c9da3}, journal = {Biol. Open}, journal-full = {Biology open}, keywords = {Centrosome;_phase_sep._/_drops}, month = Sep, pages = {1431-1440}, pmid = {27591191}, timestamp = {2017-03-02T23:22:01.000+0100}, title = {Polo kinase phosphorylation determines C. elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation}, volume = 5, year = 2016 }