An ordered pattern of Ana2 phosphorylation by Plk4 is required for centriole assembly.

The Journal of Cell Biology
Tiffany A McLamarrahGregory C Rogers

Abstract

Polo-like kinase 4 (Plk4) initiates an early step in centriole assembly by phosphorylating Ana2/STIL, a structural component of the procentriole. Here, we show that Plk4 binding to the central coiled-coil (CC) of Ana2 is a conserved event involving Polo-box 3 and a previously unidentified putative CC located adjacent to the kinase domain. Ana2 is then phosphorylated along its length. Previous studies showed that Plk4 phosphorylates the C-terminal STil/ANa2 (STAN) domain of Ana2/STIL, triggering binding and recruitment of the cartwheel protein Sas6 to the procentriole assembly site. However, the physiological relevance of N-terminal phosphorylation was unknown. We found that Plk4 first phosphorylates the extreme N terminus of Ana2, which is critical for subsequent STAN domain modification. Phosphorylation of the central region then breaks the Plk4-Ana2 interaction. This phosphorylation pattern is important for centriole assembly and integrity because replacement of endogenous Ana2 with phospho-Ana2 mutants disrupts distinct steps in Ana2 function and inhibits centriole duplication.

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Citations

Feb 26, 2019·The Journal of Biological Chemistry·Ryan A DenuMark E Burkard
Oct 3, 2019·Experimental Biology and Medicine·Shirley Jusino, Harold I Saavedra
Dec 17, 2019·The Journal of Cell Biology·Tiffany A McLamarrahGregory C Rogers
Aug 12, 2018·Cells·Anna SchwarzThomas Müller-Reichert
Aug 28, 2019·The Journal of Cell Biology·Daisuke TakaoDaiju Kitagawa
May 18, 2019·Proceedings of the National Academy of Sciences of the United States of America·Michael F PressDennis J Slamon
Jan 3, 2019·Annual Review of Biochemistry·David K Breslow, Andrew J Holland
Apr 20, 2019·Nature Communications·Shohei Yamamoto, Daiju Kitagawa
Nov 11, 2020·Open Biology·Agnieszka FatalskaDavid M Glover
Sep 22, 2020·Current Opinion in Structural Biology·Ioannis Vakonakis
Sep 22, 2020·Current Opinion in Structural Biology·Shohei Yamamoto, Daiju Kitagawa
Nov 2, 2020·Current Opinion in Structural Biology·Raksha DeviSuzanna L Prosser
Mar 25, 2021·The Journal of Cell Biology·Catarina NabaisMónica Bettencourt-Dias
Jan 9, 2022·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·Yuan TianJingyan Fu

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Methods Mentioned

BETA
transgenic
two-hybrid
coIP
pull-down
immunoprecipitation
circular dichroism
superresolution microscopy
pull-downs
Y2H
transfection

Software Mentioned

GraphPad
SIM
SR
softWoRx
Prism
Chirascan
plus

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