Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation

Oliver Wueseke, David Zwicker, Anne Schwager, Yao Liang Wong, Karen Oegema, Frank Jülicher, Anthony A. Hyman, Jeffrey B. Woodruff

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (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-54A) 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-54A 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 PLK-1 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.

Original languageEnglish (US)
Pages (from-to)1431-1440
Number of pages10
JournalBiology Open
Volume5
Issue number10
DOIs
StatePublished - Oct 15 2016
Externally publishedYes

Fingerprint

centrosomes
Centrosome
Phosphorylation
Caenorhabditis elegans
Conformations
phosphorylation
phosphotransferases (kinases)
Phosphotransferases
Scaffolds
Microtubule-Organizing Center
microtubules
Centrioles
Microtubules
mutants
centrioles
polo-like kinase 1
Theoretical Models
Embryonic Structures
Maintenance
Nucleation

Keywords

  • C. elegans
  • Centrosome
  • Microtubule
  • PCM
  • Polo-like kinase
  • SPD-5

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation. / Wueseke, Oliver; Zwicker, David; Schwager, Anne; Wong, Yao Liang; Oegema, Karen; Jülicher, Frank; Hyman, Anthony A.; Woodruff, Jeffrey B.

In: Biology Open, Vol. 5, No. 10, 15.10.2016, p. 1431-1440.

Research output: Contribution to journalArticle

Wueseke, Oliver ; Zwicker, David ; Schwager, Anne ; Wong, Yao Liang ; Oegema, Karen ; Jülicher, Frank ; Hyman, Anthony A. ; Woodruff, Jeffrey B. / Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation. In: Biology Open. 2016 ; Vol. 5, No. 10. pp. 1431-1440.
@article{6852ead11d3c4893849a7c2a0f9f239a,
title = "Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation",
abstract = "Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (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-54A) 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-54A 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 PLK-1 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.",
keywords = "C. elegans, Centrosome, Microtubule, PCM, Polo-like kinase, SPD-5",
author = "Oliver Wueseke and David Zwicker and Anne Schwager and Wong, {Yao Liang} and Karen Oegema and Frank J{\"u}licher and Hyman, {Anthony A.} and Woodruff, {Jeffrey B.}",
year = "2016",
month = "10",
day = "15",
doi = "10.1242/bio.020990",
language = "English (US)",
volume = "5",
pages = "1431--1440",
journal = "Biology Open",
issn = "2046-6390",
publisher = "Company of Biologists Ltd",
number = "10",

}

TY - JOUR

T1 - Polo-like kinase phosphorylation determines Caenorhabditis elegans centrosome size and density by biasing SPD-5 toward an assembly-competent conformation

AU - Wueseke, Oliver

AU - Zwicker, David

AU - Schwager, Anne

AU - Wong, Yao Liang

AU - Oegema, Karen

AU - Jülicher, Frank

AU - Hyman, Anthony A.

AU - Woodruff, Jeffrey B.

PY - 2016/10/15

Y1 - 2016/10/15

N2 - Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (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-54A) 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-54A 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 PLK-1 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.

AB - Centrosomes are major microtubule-organizing centers composed of centrioles surrounded by an extensive proteinacious layer called the pericentriolar material (PCM). In Caenorhabditis elegans embryos, the mitotic PCM expands by Polo-like kinase 1 (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-54A) 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-54A 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 PLK-1 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.

KW - C. elegans

KW - Centrosome

KW - Microtubule

KW - PCM

KW - Polo-like kinase

KW - SPD-5

UR - http://www.scopus.com/inward/record.url?scp=84994017882&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84994017882&partnerID=8YFLogxK

U2 - 10.1242/bio.020990

DO - 10.1242/bio.020990

M3 - Article

C2 - 27591191

AN - SCOPUS:84994017882

VL - 5

SP - 1431

EP - 1440

JO - Biology Open

JF - Biology Open

SN - 2046-6390

IS - 10

ER -