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Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii

PLoS Pathogens, ISSN: 1553-7366, Vol: 4, Issue: 2, Page: e36
2008
  • 82
    Citations
  • 0
    Usage
  • 87
    Captures
  • 0
    Mentions
  • 31
    Social Media
Metric Options:   Counts1 Year3 Year

Metrics Details

  • Citations
    82
  • Captures
    87
  • Social Media
    31
    • Shares, Likes & Comments
      31
      • Facebook
        31

Article Description

Apicomplexa are obligate intracellular pathogens that have fine-tuned their proliferative strategies to match a large variety of host cells. A critical aspect of this adaptation is a flexible cell cycle that remains poorly understood at the mechanistic level. Here we describe a forward genetic dissection of the apicomplexan cell cycle using the Toxoplasma model. By high-throughput screening, we have isolated 165 temperature sensitive parasite growth mutants. Phenotypic analysis of these mutants suggests regulated progression through the parasite cell cycle with defined phases and checkpoints. These analyses also highlight the critical importance of the peculiar intranuclear spindle as the physical hub of cell cycle regulation. To link these phenotypes to parasite genes, we have developed a robust complementation system based on a genomic cosmid library. Using this approach, we have so far complemented 22 temperature sensitive mutants and identified 18 candidate loci, eight of which were independently confirmed using a set of sequenced and arrayed cosmids. For three of these loci we have identified the mutant allele. The genes identified include regulators of spindle formation, nuclear trafficking, and protein degradation. The genetic approach described here should be widely applicable to numerous essential aspects of parasite biology. © 2008 Gubbels et al.

Bibliographic Details

Marc-Jan Gubbels; Margaret Lehmann; Mani Muthalagi; Maria E Jerome; Carrie F Brooks; Tomasz Szatanek; Jayme Flynn; Ben Parrot; Josh Radke; Boris Striepen; Michael W White; Kami Kim

Public Library of Science (PLoS)

Immunology and Microbiology; Biochemistry, Genetics and Molecular Biology

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