Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii.
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 disse...
| Main Authors: | , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2008-02-01
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| Series: | PLoS Pathogens |
| Online Access: | http://europepmc.org/articles/PMC2242837?pdf=render |
| _version_ | 1798045429092319232 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Marc-Jan Gubbels |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-11T23:21:58Z |
| format | Article |
| id | doaj.art-484811f118a2477ba1bd8a0ead8273e0 |
| institution | Directory Open Access Journal |
| issn | 1553-7366 1553-7374 |
| language | English |
| last_indexed | 2024-04-11T23:21:58Z |
| publishDate | 2008-02-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj.art-484811f118a2477ba1bd8a0ead8273e02022-12-22T03:57:26ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742008-02-0142e3610.1371/journal.ppat.0040036Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii.Marc-Jan GubbelsMargaret LehmannMani MuthalagiMaria E JeromeCarrie F BrooksTomasz SzatanekJayme FlynnBen ParrotJosh RadkeBoris StriepenMichael W WhiteApicomplexa 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.http://europepmc.org/articles/PMC2242837?pdf=render |
| spellingShingle | 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 Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. PLoS Pathogens |
| title | Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. |
| title_full | Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. |
| title_fullStr | Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. |
| title_full_unstemmed | Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. |
| title_short | Forward genetic analysis of the apicomplexan cell division cycle in Toxoplasma gondii. |
| title_sort | forward genetic analysis of the apicomplexan cell division cycle in toxoplasma gondii |
| url | http://europepmc.org/articles/PMC2242837?pdf=render |
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