Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Accepted manuscripts
    • 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
    • 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
Genetic and morphological evidence for two parallel pathways of cell-cycle-coupled cytokinesis in Dictyostelium
Akira Nagasaki, Eugenio L. de Hostos, Taro Q. P. Uyeda
Journal of Cell Science 2002 115: 2241-2251;
Akira Nagasaki
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Eugenio L. de Hostos
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Taro Q. P. Uyeda
  • 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. 2.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 2.

    Disruption constructs targeting mhcA and amiA, and confirmation of disruption using genomic PCR. (A) The targeting vector used to knockout amiA was constructed by replacing part of its coding region and promoter with the blasticidin S [pKO amiA(Bsr)] or G418 [pKO amiA(Neo)] resistance gene. In the targeting vector for mhcA, a portion of the motor domain was replaced with the G418 resistance cassette. Expression of all drug resistance genes was driven by the actin 15 promoter. Thick lines indicate coding sequences of amiA and mhcA. (B) Mutant cells were identified by a shift in size of the PCR products. (Left) Knockout of amiA in mhcA- cells (HS1), yielding HTU1; (middle) knockout of amiA in corA- cells, yielding HTU8; (right) knockout of corA in mhcA- cells (HS1), yielding HTU7. Arrows in A show the positions of the primers used for genomic PCR.

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

    Three modes of cytokinesis in Dictyostelium. A series of time-lapse phase contrast images obtained at 30 second intervals are shown. (A) Wild-type AX2 cells were embedded in low melting temperature agarose and cultured without solid surfaces. In this condition, wild-type cells divide using cytokinesis A. (B) mhcA- cells were cultured on a plastic dish to allow adhesion to a solid surface. Division of mhcA- cells is driven by cytokinesis B. (C) Multinucleate mhcA- cells grown in suspension for 3 days were then placed on a plastic dish. These giant cells divide by cytokinesis C. Bars, 10μ m.

  • Table 1.

    Strains used in this study

    GenotypeParental strainSource
    AX2— Watts and Ashworth, 1970
    HTU1 amiA (Bsr) AX2 Nagasaki et al., 1998
    HS1 mhcA (Thy) AX3 Ruppel et al., 1994
    coronin- corA (Bsr) AX2 Fukui et al., 1999
    HTU2 amiA (Neo) AX2This study
    HTU3 mhcA (Neo) AX2This study
    HTU4 amiA (Bsr), mhcA (Thy) AX3This study
    HTU5 amiA (Bsr), mhcA (Neo) AX2This study
    HTU6 amiA (Neo), mhcA (Thy) AX3This study
    HTU7 corA (Bsr), mhcA (Neo) AX2This study
    HTU8 amiA (Neo), corA (Bsr) AX2This study
    Two strains of mhcA- and three strains of amiA-lmhcA- cells were created to assess possible phenotypic differences derived from differences in the parent strains. These redundant strains showed identical phenotypes in our assays, however, and results from HTU3, HTU5 and HTU6 are not presented. The HS1 mhcA- strain was originally constructed by Ruppel et al. ( Ruppel et al., 1994) using the THY1 gene (Dynes and Firtel, 1989) as a selection marker in JH10, a thymidine auxotroph derivative of AX3 (Hadwiger and Firtel, 1992).
  •   Fig. 3.
    • Download figure
    • Open in new tab
    • Download powerpoint
    Fig. 3.

    Phenotypic analysis of wild-type and mutant cells in which cytokinesis A and/or B has been disrupted. (A,B) Histograms showing the distributions of nuclei/cell among cells grown in suspension (B) or on glass surfaces (A). Cells of all three strains shown were cultured in suspension before being transferred to plastic Petri dishes with thin glass bottoms; fixation, DAPI staining and counting of nuclei were performed immediately (B) or after 3 days of continued growth on the glass surface (A). (C-I) Photomicrographs of cells grown for 3 days on a glass surface. Unlike cells shown in A and B, these cells were precultured on plastic Petri dishes, suspended by streams of medium using micropipettes, diluted, and replated to plastic Petri dishes with thin glass bottoms because cells carrying the mhcA- mutation were unable to grow in suspension. Wild-type (C) and mhcA- cells (D) were mostly mononucleate. AmiA- (E), corA- (G) and amiA-/corA- (I) cells were somewhat larger and flatter than wild-type cells, indicating moderate disruption of cytokinesis. The greater enlargement of amiA-/mhcA- (F) and corA-/mhcA- (H) double knockout cells suggests cytokinesis is more severely affected in these strains. (J) A histogram showing the distributions of nuclei/cell among the cells shown in C-I. More than half of the amiA-/mhcA- and corA-/mhcA- cells are multinucleate, and more than 30% of them are highly multinucleate (more than five nuclei per cell), confirming severe disruption of cytokinesis in these cells. Magnifications in panels C-I are the same. Bar, 10 μm.

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

    Sequences of morphological changes during division on a solid substrate. Cells were attached to plastic dishes for 2 days before observation. Each panel shows a series of phase contrast images recorded with intervals of 60 seconds between frames. (A-E) Mitotic wild-type (A), mitotic mhcA- (B), mitotic amiA- (C), mitotic corA- (D) and amiA-/mhcA- (E) cells. The cell cycle stage of the cell in E was not determined, but is most probably interphase. Bars, 10 μm. Magnifications of A-D are the same.

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

    Visualization of the cell cycle as a function of GFP-histone expression. (A) Structure of the GFP-tagged histone H1 chimeric gene. (B,C) Confocal images of the nucleus were identified in interphase (B) and mitotic (C) cells by GFP-H1 fluorescence. (D-F) A series of time-lapse micrographs of GFP fluorescence images superimposed on phase contrast images. They show differing cytokinesis processes in three strains of GFP-H1-expressing cells grown on solid substrates: mhcA- cells undergoing cytokinesis B (D); amiA- cells failing to divide (E); and amiA-/mhcA- cells undergoing cytokinesis C (F). Bars, 10 μm.

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

    A schematic diagram depicting two pathways leading to cell-cycle-coupled cell division in Dictyostelium cells. (A) Cytokinesis A. Mitotic AmiA- or coronin- cell carries out cytokinesis by active contraction of the cleavage furrow which depends on actin and myosin II. (B) Cytokinesis B. A mitotic myosin II-null cell divides by passive contraction of the cleavage furrow. In this case, cytoplasm in equatorial region is withdrawn indirectly (white arrows inside the cell) by traction forces generated along polar peripheries (black arrows). (C) Summary of three methods of cytokinesis in Dictyostelium. Cytokinesis A requires myosin II expression, but adhesion to a substrate is not necessary. Cytokinesis B is not dependent on myosin II but adhesion is indispensable. These two mechanisms of cell division occur immediately following nuclear division and are somehow coordinated in wild-type cells. The third pathway, cytokinesis C, is cell cycle independent and occurs during interphase.

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.
Genetic and morphological evidence for two parallel pathways of cell-cycle-coupled cytokinesis in Dictyostelium
(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
Genetic and morphological evidence for two parallel pathways of cell-cycle-coupled cytokinesis in Dictyostelium
Akira Nagasaki, Eugenio L. de Hostos, Taro Q. P. Uyeda
Journal of Cell Science 2002 115: 2241-2251;
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
Citation Tools
Research Article
Genetic and morphological evidence for two parallel pathways of cell-cycle-coupled cytokinesis in Dictyostelium
Akira Nagasaki, Eugenio L. de Hostos, Taro Q. P. Uyeda
Journal of Cell Science 2002 115: 2241-2251;

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
    • Materials and Methods
    • Results
    • Discussion
    • Acknowledgments
    • Footnotes
    • References
  • Figures & tables
  • Supp info
  • Info & metrics
  • PDF

Related articles

Cited by...

More in this TOC section

  • Switching between blebbing and lamellipodia depends on the degree of non-muscle myosin II activity
  • Kindlin-2 promotes rear focal adhesion disassembly and directional persistence during cell migration
  • Histone chaperone APLF level dictates the implantation of mouse embryos
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
  • 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