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
The Ste20-like kinase SvkA of Dictyostelium discoideum is essential for late stages of cytokinesis
Meino Rohlfs, Rajesh Arasada, Petros Batsios, Julia Janzen, Michael Schleicher
Journal of Cell Science 2007 120: 4345-4354; doi: 10.1242/jcs.012179
Meino Rohlfs
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Rajesh Arasada
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Petros Batsios
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Julia Janzen
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Michael Schleicher
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF
Loading

Article Figures & Tables

Figures

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

    Disruption of the svkA gene encoding severin kinase. (A) Two fragments of the kinase were amplified by PCR from genomic DNA, ligated to the blasticidin-resistance cassette and electroporated into AX2 wild-type cells. (B) Blasticidin-resistant clones were tested for knockout events by PCR1 (primers 1 and 2, 1385 bp for knockout) and PCR2 (primers 1 and 3, 2038 bp as a wild-type control). Under the conditions used, PCR2 did not ever amplify ∼3400 bp, including the blasticidin-resistance cassette, in the knockout, but the lack of a PCR product indicated a disruption of the svkA gene as well. (C) Western blot of 2×105 cells per lane, visualized with kinase-specific antibodies confirms the knockout at the level of SvkA protein and also shows the expression of the GFP fusion protein in the wild-type (first lane) and the mutant background (fourth lane). (D) GFP-SvkA-FL, immunoprecipitated from rescue cells, phosphorylated domains 2 and 3 of severin (DS211C, left), myelin basic protein (MyBP, right) and itself (both lanes).

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

    Expression of SvkA during development and induction of cytofission. (A) Western blot with antibodies against SvkA showing the presence of the kinase throughout the 24 hours of wild-type development (2×105 cells per lane). (B) The knockout cells grown on a plastic surface were considerably larger than the wild-type cells and the rescue cells. (C,D) SvkA-knockout cells were shifted from HL-5 medium to low-salt phosphate buffer (PB) and observed for several hours. The large and multinucleated cells separated only partially. Arrows indicate persistent connections between cells. Bars, 50 μm (B,C); 10 μm (D); see also supplementary material Movie 1.

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

    Developmental defects in the svkA-null mutant. (A) The development of starved svkA-null cells on phosphate agar showed a delay, whereas the wild-type and rescue strains developed as expected into loose and tight aggregates (6 hours/9 hours), slugs (12 hours) and mature fruiting bodies (24 hours). (B) Typical fruiting bodies obtained after three days on phosphate agar (wild-type and rescue strains: one each at the right; knockout cells: nine examples on the left). (C) Phototaxis of D. discoideum slugs on phosphate agar towards a light source at the right (*).

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

    Defective cytokinesis in svkA-null cells. (A,B) Cells grown on a solid surface or in a shaking culture (150 rpm) were fixed and stained with DAPI for quantification of nuclei per cell. More than 1000 cells were counted for each cell line. In the mutant, >50% of the total number of nuclei were found in multinucleated cells under both conditions.

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

    Asymmetric cytokinesis. (A,B) Time-lapse images of svkA-knockout cells dividing on a plastic surface. The increasingly asymmetric cleavage furrow inhibits complete separation of the daughter cells (bars, 20 μm). (C) Time-lapse series of projections calculated from stacks of confocal images with GFP-cortexillin I expressed in svkA-knockout cells. The dividing cell shows an asymmetric cleavage furrow and a remaining cytoplasmic bridge that eventually ruptures ∼8 minutes after the onset of cytokinesis (bar, 5 μm).

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

    Analysis of cytokinesis with marker proteins. (A-G) Confocal sections of fixed svkA-knockout cells representing different stages and the symmetrical population of SvkA-null cell division. All cells are stained for alpha-tubulin and in addition for F-actin (A,B) (the arrow in A indicates the dividing cell), cortexillin I (C,E), myosin II (D), DGAP1 (F) or coronin (G). Bars, 5 μm.

  •   Fig. 7.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 7.

    Enrichment of SvkA around the centrosome, the spindle and the midzone. (A) SvkA-knockout cells expressing GFP-SvkA-FL were fixed and stained with antibodies against alpha-tubulin. Three confocal sections are shown. GFP-SvkA-FL is enriched diffusely around the centrosome of interphase cells (open arrow) and around the spindle in mitotic cells (closed arrows). (B) Confocal section of fixed wild-type cells expressing the C-terminal domain construct GFP-SvkA-CT. The arrow indicates the localization of GFP-SvkA-CT around the centrosome. (C) A confocal section of fixed wild-type cells expressing GFP-SvkA-FL shows colocalization with the DNA (upper panel) and, at later stages, around the spindle (lower panel). Bars, 5 μm.

  •   Fig. 8.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 8.

    Live-cell recordings show SvkA in the midzone. (A) Wild-type cells expressing GFP-SvkA-FL accumulate the kinase only after the appearance of the cleavage furrow. The arrows indicate the localization of SvkA in the midzone during cell separation. (B) A svkA-knockout cell (confocal section of live cell, agar overlay) containing four nuclei (stars) and expressing the CT-terminal domain of SvkA. GFP-SvkA-CT determines localization to the midzone but is not sufficient to prevent incomplete cell division (see renewed fusion of partially separated cells in frame 11′). (C) SvkA-knockout cells expressing the dead kinase show that GFP-SvkA-K134A-FL localizes to the late cleavage furrow (arrows). (The stars indicate the approximate localization of the nuclei outside the focus plane. Bars, 5 μm.)

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.
The Ste20-like kinase SvkA of Dictyostelium discoideum is essential for late stages of cytokinesis
(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
The Ste20-like kinase SvkA of Dictyostelium discoideum is essential for late stages of cytokinesis
Meino Rohlfs, Rajesh Arasada, Petros Batsios, Julia Janzen, Michael Schleicher
Journal of Cell Science 2007 120: 4345-4354; doi: 10.1242/jcs.012179
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Research Article
The Ste20-like kinase SvkA of Dictyostelium discoideum is essential for late stages of cytokinesis
Meino Rohlfs, Rajesh Arasada, Petros Batsios, Julia Janzen, Michael Schleicher
Journal of Cell Science 2007 120: 4345-4354; doi: 10.1242/jcs.012179

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
    • Acknowledgments
    • Footnotes
    • References
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF

Related articles

Cited by...

More in this TOC section

  • Hydrostatic pressure prevents chondrocyte differentiation through heterochromatin remodeling
  • PLEKHG4B enables actin cytoskeletal remodeling during epithelial cell–cell junction formation
  • Matrix stiffness regulates α-TAT1-mediated acetylation of α-tubulin and promotes silica-induced epithelial–mesenchymal transition via DNA damage
Show more RESEARCH ARTICLE

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