Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

User menu

  • Log in

Search

  • Advanced search
Journal of Cell Science
  • COB
    • About The Company of Biologists
    • Development
    • Journal of Cell Science
    • Journal of Experimental Biology
    • Disease Models & Mechanisms
    • Biology Open

supporting biologistsinspiring biology

Journal of Cell Science

  • Log in
Advanced search

RSS   Twitter  Facebook   YouTube  

  • Home
  • Articles
    • Accepted manuscripts
    • Issue in progress
    • Latest complete issue
    • Issue archive
    • Archive by article type
    • Special issues
    • Subject collections
    • Cell Scientists to Watch
    • First Person
    • Sign up for alerts
  • About us
    • About JCS
    • Editors and Board
    • Editor biographies
    • Travelling Fellowships
    • Grants and funding
    • Journal Meetings
    • Workshops
    • The Company of Biologists
    • Journal news
  • For authors
    • Submit a manuscript
    • Aims and scope
    • Presubmission enquiries
    • Fast-track manuscripts
    • Article types
    • Manuscript preparation
    • Cover suggestions
    • Editorial process
    • Promoting your paper
    • Open Access
    • JCS Prize
    • Manuscript transfer network
    • Biology Open transfer
  • Journal info
    • Journal policies
    • Rights and permissions
    • Media policies
    • Reviewer guide
    • Sign up for alerts
  • Contacts
    • Contact JCS
    • Subscriptions
    • Advertising
    • Feedback
Research Article
Cell-cycle-regulated expression of STIL controls centriole number in human cells
Christian Arquint, Katharina F. Sonnen, York-Dieter Stierhof, Erich A. Nigg
Journal of Cell Science 2012 125: 1342-1352; doi: 10.1242/jcs.099887
Christian Arquint
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Katharina F. Sonnen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
York-Dieter Stierhof
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Erich A. Nigg
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • For correspondence: erich.nigg@unibas.ch
  • Article
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF + SI
  • PDF
Loading

Article Figures & Tables

Figures

  • Fig. 1.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 1.

    STIL is required for centriole duplication in U2OS cells. Asynchronously growing U2OS cells were transfected for 72 hours with control siRNA (GL2), three different STIL siRNAs (STIL1–STIL3) or PLK4 siRNA oligonucleotides. (A) Western blot analysis of STIL protein levels in control (GL2) and STIL-depleted U2OS cell lysates (STIL1–STIL3), using the STIL antibody ab89314 and an antibody against α-tubulin as a loading control. (B) After siRNA treatment, U2OS cells were fixed and stained with antibodies against CP110 (red) and Cep135 (green) for immunofluorescence microscopy. DNA was stained with DAPI (blue). Yellow rectangles illustrate probable orientation of centrioles. (C) Centriole numbers per cell in response to treatment with GL2, STIL or PLK4 siRNA, as in B. (n=3, 100 cells were analyzed in each experiment, error bars denote s.d.). Scale bars: 1 μm (magnifications) and 5 μm (overview; Merge + DAPI).

  • Fig. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    STIL specifically localizes to the proximal end of daughter centrioles. (A) U2OS cells fixed and stained with antibodies against STIL (ca66, green) and CP110 (red); DNA (DAPI) is shown in gray. (B) STIL localizes closer to Cep164-negative centrioles. U2OS cells fixed and stained with antibodies against STIL (ca66, green), CP110 (red) and Cep164 (blue); DNA (DAPI) is shown in gray. The presumed orientation of the Cep164-positive mother centriole (M) is depicted with a yellow rectangle, the orientation of the respective Cep164-negative daughter centriole (D) with a white rectangle. The scheme to the left illustrates the relative localization of STIL (green), CP110 (red) and Cep164 (blue) within this centriole pair. (C) STIL colocalizes with SAS-6. U2OS cells were fixed and stained with antibodies against STIL (ca66, green), SAS-6 (red) and CP110 (blue); DNA (DAPI) is shown in gray. The presumed orientation of the SAS-6-negative mother centriole (M) is depicted with a yellow rectangle, the orientation of the respective SAS-6-positive daughter centriole (D) with a white rectangle. The scheme to the left illustrates the relative localization of STIL (green), SAS-6 (red) and CP110 (blue) within this centriole pair. (D) As shown by immunoelectron microscopy STIL localizes close to the interphase between mother and daughter centrioles (top row). U2OS cells were fixed and incubated with antibodies against STIL (ca66), followed by gold-labeled secondary antibodies. Schematic representations (bottom row) illustrate the orientation of mother (M) and daughter (D) centrioles; the localization of gold particles is denoted by black dots. Scale bars: 5 μm (A–C, Merge + DAPI) and 1 μm (A–C, all higher magnifications), 0.5 μm (D).

  • Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Localization of STIL to centrioles is cell-cycle dependent. (A) Asynchronously growing U2OS cells fixed and stained for immunofluorescence microscopy with antibodies against STIL (ca66, green), CP110 (red) and Cep135 (blue); DNA (DAPI) is shown in gray. Representative images are shown for the different cell cycle stages (determined by the number of centrioles in interphase and DNA stain in mitosis). (B) Nocodazole (Noco)-arrested U2OS cells were released into mitosis in the presence or absence of MG132 and subjected to western blot analysis at different time points after release, using antibodies against STIL (ab89314) and cyclin B1. α-tubulin was monitored as loading control. (C) Same experiment as described in B except that Cdk1 inhibitor (RO-3306) was added to both MG132-treated and untreated cells. Anti-phospho-histone H3 (Ser10) antibodies were used to monitor mitotic progression. (D) Nocodazole (Noco)-arrested HeLa S3 cells previously treated with GL2 (control) or siRNA oligonucleotides targeting Cdc20 and/or Cdh1 were released into mitosis in the presence of Cdk1 inhibitor (RO-3306) and subjected to western blot analysis at different time points after release, using antibodies against STIL (ab89314), cyclin B1, Cdc20 and Cdh1. α-tubulin was monitored as a loading control. Scale bars: 5 μm (Merge + DAPI) and 1 μm (higher magnifications).

  • Fig. 4.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 4.

    Relationship between STIL, Ana2 and SAS-5. (A) Phylogenetic tree illustrating relationship between metazoan STIL and Drosophila Ana2 protein families. The tree was generated on phylogeny.fr (www.phylogeny.fr) using MUSCLE for alignment of amino acid sequences and PhyML (maximum likelihood method) for phylogeny. Number of amino acids (aa) for each sequence are indicated to the right. The scale bar indicates 0.5 substitutions per site. (B) Multiple protein sequence alignment (MUSCLE; colored according to the BLOSUM62 score, conservation visibility value was set to 30) of STIL/Ana2-related protein sequences. Regions encompassing highest sequence conservation (colored boxes on a scheme of human STIL) are shown. Alignment of three SAS-5 protein sequences of nematodes is shown below. STAN, STIL/ANA2; TIM, truncated in microcephaly.

  • Fig. 5.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 5.

    SAS-6 localizes to centrioles in the absence of STIL. (A) Asynchronously growing U2OS cells transfected for 72 hours with GL2, STIL or SAS-6 siRNA oligonucleotides. Cells were fixed and stained with antibodies against STIL (ca66, green), SAS-6 (red) and CP110 (blue) for immunofluorescence microscopy; DNA (DAPI) is depicted in gray. Representative images of prophase cells are shown (determined by DAPI staining). (B) Quantification of centriolar levels of STIL and SAS-6 fluorescence in cells treated as described in A. Only prophase cells (determined by DAPI staining) were considered. (n=3, 10 centriolar pairs were analyzed in each experiment, error bars denote s.d.). Note that in SAS-6-depleted cells, STIL is completely absent from centrioles, whereas in STIL-depleted cells, SAS-6 localizes to centrioles (albeit in reduced amounts). (C) Western blot analysis of STIL and SAS-6 protein levels in control- (GL2), STIL- (STIL 1-3) and SAS-6-depleted U2OS cell lysates, probed with antibodies against STIL (ab89314), SAS-6 and α-tubulin for loading control. Scale bars: 5 μm (Merge + DAPI) and 1 μm (higher magnifications).

  • Fig. 6.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 6.

    STIL overexpression results in centriole amplification. Asynchronously growing U2OS cells were transfected with plasmids encoding FLAG-tagged STIL, PLK4 or SAS-6 or empty vector. After 48 hours, cells were fixed and stained with antibodies against the FLAG tag (green), CP110 (red) and Cep135 (blue); DNA (DAPI) is shown in gray. Cells were then analyzed by fluorescence microscopy. (A) The number of centrioles per transfected cell was determined based on CP110 staining. The graph illustrates the percentage of cells in each indicated category for control-, STIL- PLK4- and SAS-6-transfected cells (n=3, 50 cells were analyzed in each experiment, error bars denote s.d.). (B) Representative images are shown for STIL-transfected and control cells. Note that in cells overexpressing STIL, multiple procentrioles form around a single centriole. (C) U2OS cells transiently expressing FLAG-tagged STIL were further stained with antibodies against CP110 (red), STIL (ca66, green) and Cep164 (blue). Representative images are shown and a scheme illustrates the localization of CP110 (red), STIL (green) and Cep164 (blue) to the flower-like structures. Note that Cep164, a marker for mature mother centrioles, localizes to the central centriole.(D) U2OS cells transiently expressing FLAG-tagged STIL were stained with antibodies against CP110 (red), STIL (ca66, green) and SAS-6 (blue). Note that both STIL and SAS-6 colocalize as a ring around the central centriole. Scale bars: 5 μm (Merge + DAPI) and 1 μm (all higher magnifications).

Previous ArticleNext Article
Back to top
Previous ArticleNext Article

This Issue

 Download PDF

Email

Thank you for your interest in spreading the word on Journal of Cell Science.

NOTE: We only request your email address so that the person you are recommending the page to knows that you wanted them to see it, and that it is not junk mail. We do not capture any email address.

Enter multiple addresses on separate lines or separate them with commas.
Cell-cycle-regulated expression of STIL controls centriole number in human cells
(Your Name) has sent you a message from Journal of Cell Science
(Your Name) thought you would like to see the Journal of Cell Science web site.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Share
Research Article
Cell-cycle-regulated expression of STIL controls centriole number in human cells
Christian Arquint, Katharina F. Sonnen, York-Dieter Stierhof, Erich A. Nigg
Journal of Cell Science 2012 125: 1342-1352; doi: 10.1242/jcs.099887
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Research Article
Cell-cycle-regulated expression of STIL controls centriole number in human cells
Christian Arquint, Katharina F. Sonnen, York-Dieter Stierhof, Erich A. Nigg
Journal of Cell Science 2012 125: 1342-1352; doi: 10.1242/jcs.099887

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Alerts

Please log in to add an alert for this article.

Sign in to email alerts with your email address

Article navigation

  • Top
  • Article
    • Summary
    • Introduction
    • Results
    • Discussion
    • Materials and Methods
    • Acknowledgements
    • Footnotes
    • References
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF + SI
  • PDF

Related articles

Cited by...

More in this TOC section

  • EFA6A, an exchange factor for Arf6, regulates early steps in ciliogenesis
  • SetDB1 and Su(var)3-9 play non-overlapping roles in somatic cell chromosomes of Drosophila melanogaster
  • p.E152K-STIM1 mutation deregulates Ca2+ signaling contributing to chronic pancreatitis
Show more Research Articles

Similar articles

Other journals from The Company of Biologists

Development

Journal of Experimental Biology

Disease Models & Mechanisms

Biology Open

Advertisement

2020 at The Company of Biologists

Despite the challenges of 2020, we were able to bring a number of long-term projects and new ventures to fruition. While we look forward to a new year, join us as we reflect on the triumphs of the last 12 months.


Mole – The Corona Files

"This is not going to go away, 'like a miracle.' We have to do magic. And I know we can."

Mole continues to offer his wise words to researchers on how to manage during the COVID-19 pandemic.


Cell scientist to watch – Christine Faulkner

In an interview, Christine Faulkner talks about where her interest in plant science began, how she found the transition between Australia and the UK, and shares her thoughts on virtual conferences.


Read & Publish participation extends worldwide

“The clear advantages are rapid and efficient exposure and easy access to my article around the world. I believe it is great to have this publishing option in fast-growing fields in biomedical research.”

Dr Jaceques Behmoaras (Imperial College London) shares his experience of publishing Open Access as part of our growing Read & Publish initiative. We now have over 60 institutions in 12 countries taking part – find out more and view our full list of participating institutions.


JCS and COVID-19

For more information on measures Journal of Cell Science is taking to support the community during the COVID-19 pandemic, please see here.

If you have any questions or concerns, please do not hestiate to contact the Editorial Office.

Articles

  • Accepted manuscripts
  • Issue in progress
  • Latest complete issue
  • Issue archive
  • Archive by article type
  • Special issues
  • Subject collections
  • Interviews
  • Sign up for alerts

About us

  • About Journal of Cell Science
  • Editors and Board
  • Editor biographies
  • Travelling Fellowships
  • Grants and funding
  • Journal Meetings
  • Workshops
  • The Company of Biologists

For Authors

  • Submit a manuscript
  • Aims and scope
  • Presubmission enquiries
  • Fast-track manuscripts
  • Article types
  • Manuscript preparation
  • Cover suggestions
  • Editorial process
  • Promoting your paper
  • Open Access
  • JCS Prize
  • Manuscript transfer network
  • Biology Open transfer

Journal Info

  • Journal policies
  • Rights and permissions
  • Media policies
  • Reviewer guide
  • Sign up for alerts

Contacts

  • Contact JCS
  • Subscriptions
  • Advertising
  • Feedback

Twitter   YouTube   LinkedIn

© 2021   The Company of Biologists Ltd   Registered Charity 277992