A nuclear targeting system in <it>Plasmodium falciparum</it>
<p>Abstract</p> <p>Background</p> <p>The distinct differences in gene control mechanisms acting in the nucleus between <it>Plasmodium falciparum </it>and the human host could lead to new potential drug targets for anti-malarial development. New molecular too...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
BMC
2010-05-01
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| Series: | Malaria Journal |
| Online Access: | http://www.malariajournal.com/content/9/1/126 |
| _version_ | 1818229581550714880 |
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| author | Kochakarn Theerarat Songprakhon Pucharee Tinikul Ruchanok Kumpornsin Krittikorn Uthaipibull Chairat Wittayacom Kanjana Chookajorn Thanat |
| author_facet | Kochakarn Theerarat Songprakhon Pucharee Tinikul Ruchanok Kumpornsin Krittikorn Uthaipibull Chairat Wittayacom Kanjana Chookajorn Thanat |
| author_sort | Kochakarn Theerarat |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>The distinct differences in gene control mechanisms acting in the nucleus between <it>Plasmodium falciparum </it>and the human host could lead to new potential drug targets for anti-malarial development. New molecular toolkits are required for dissecting molecular machineries in the <it>P. falciparum </it>nucleus. One valuable tool commonly used in model organisms is protein targeting to specific sub-cellular locations. Targeting proteins to specified locations allows labeling of organelles for microscopy, or testing of how the protein of interest modulates organelle function. In recent years, this approach has been developed for various malaria organelles, such as the mitochondrion and the apicoplast. A tool for targeting a protein of choice to the <it>P. falciparum </it>nucleus using an exogenous nuclear localization sequence is reported here.</p> <p>Methods</p> <p>To develop a nuclear targeting system, a putative nuclear localization sequence was fused with green fluorescent protein (GFP). The nuclear localization sequence from the yeast transcription factor Gal4 was chosen because of its well-defined nuclear localization signal. A series of truncated Gal4 constructs was also created to narrow down the nuclear localization sequence necessary for <it>P. falciparum </it>nuclear import. Transfected parasites were analysed by fluorescent and laser-scanning confocal microscopy.</p> <p>Results</p> <p>The nuclear localization sequence of Gal4 is functional in <it>P. falciparum</it>. It effectively transported GFP into the nucleus, and the first 74 amino acid residues were sufficient for nuclear localization.</p> <p>Conclusions</p> <p>The Gal4 fusion technique enables specific transport of a protein of choice into the <it>P. falciparum </it>nucleus, and thus provides a tool for labeling nuclei without using DNA-staining dyes. The finding also indicates similarities between the nuclear transport mechanisms of yeast and <it>P. falciparum</it>. Since the nuclear transport system has been thoroughly studied in yeast, this could give clues to research on the same mechanism in <it>P. falciparum</it>.</p> |
| first_indexed | 2024-12-12T10:20:53Z |
| format | Article |
| id | doaj.art-ed0179a325b44af2beb8743e7b576981 |
| institution | Directory Open Access Journal |
| issn | 1475-2875 |
| language | English |
| last_indexed | 2024-12-12T10:20:53Z |
| publishDate | 2010-05-01 |
| publisher | BMC |
| record_format | Article |
| series | Malaria Journal |
| spelling | doaj.art-ed0179a325b44af2beb8743e7b5769812022-12-22T00:27:34ZengBMCMalaria Journal1475-28752010-05-019112610.1186/1475-2875-9-126A nuclear targeting system in <it>Plasmodium falciparum</it>Kochakarn TheeraratSongprakhon PuchareeTinikul RuchanokKumpornsin KrittikornUthaipibull ChairatWittayacom KanjanaChookajorn Thanat<p>Abstract</p> <p>Background</p> <p>The distinct differences in gene control mechanisms acting in the nucleus between <it>Plasmodium falciparum </it>and the human host could lead to new potential drug targets for anti-malarial development. New molecular toolkits are required for dissecting molecular machineries in the <it>P. falciparum </it>nucleus. One valuable tool commonly used in model organisms is protein targeting to specific sub-cellular locations. Targeting proteins to specified locations allows labeling of organelles for microscopy, or testing of how the protein of interest modulates organelle function. In recent years, this approach has been developed for various malaria organelles, such as the mitochondrion and the apicoplast. A tool for targeting a protein of choice to the <it>P. falciparum </it>nucleus using an exogenous nuclear localization sequence is reported here.</p> <p>Methods</p> <p>To develop a nuclear targeting system, a putative nuclear localization sequence was fused with green fluorescent protein (GFP). The nuclear localization sequence from the yeast transcription factor Gal4 was chosen because of its well-defined nuclear localization signal. A series of truncated Gal4 constructs was also created to narrow down the nuclear localization sequence necessary for <it>P. falciparum </it>nuclear import. Transfected parasites were analysed by fluorescent and laser-scanning confocal microscopy.</p> <p>Results</p> <p>The nuclear localization sequence of Gal4 is functional in <it>P. falciparum</it>. It effectively transported GFP into the nucleus, and the first 74 amino acid residues were sufficient for nuclear localization.</p> <p>Conclusions</p> <p>The Gal4 fusion technique enables specific transport of a protein of choice into the <it>P. falciparum </it>nucleus, and thus provides a tool for labeling nuclei without using DNA-staining dyes. The finding also indicates similarities between the nuclear transport mechanisms of yeast and <it>P. falciparum</it>. Since the nuclear transport system has been thoroughly studied in yeast, this could give clues to research on the same mechanism in <it>P. falciparum</it>.</p>http://www.malariajournal.com/content/9/1/126 |
| spellingShingle | Kochakarn Theerarat Songprakhon Pucharee Tinikul Ruchanok Kumpornsin Krittikorn Uthaipibull Chairat Wittayacom Kanjana Chookajorn Thanat A nuclear targeting system in <it>Plasmodium falciparum</it> Malaria Journal |
| title | A nuclear targeting system in <it>Plasmodium falciparum</it> |
| title_full | A nuclear targeting system in <it>Plasmodium falciparum</it> |
| title_fullStr | A nuclear targeting system in <it>Plasmodium falciparum</it> |
| title_full_unstemmed | A nuclear targeting system in <it>Plasmodium falciparum</it> |
| title_short | A nuclear targeting system in <it>Plasmodium falciparum</it> |
| title_sort | nuclear targeting system in it plasmodium falciparum it |
| url | http://www.malariajournal.com/content/9/1/126 |
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