Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system.
The African trypanosome, Trypanosoma brucei possesses a large and unique intraflagellar structure called the paraflagellar rod (PFR). The PFR is composed of 2 major proteins, PFRA and PFRC. We have generated an inducible mutant trypanosome cell line (snl-2) that expresses linked inverted copies of a...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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2000
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| _version_ | 1826257281583415296 |
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| author | Bastin, P Ellis, K Kohl, L Gull, K |
| author_facet | Bastin, P Ellis, K Kohl, L Gull, K |
| author_sort | Bastin, P |
| collection | OXFORD |
| description | The African trypanosome, Trypanosoma brucei possesses a large and unique intraflagellar structure called the paraflagellar rod (PFR). The PFR is composed of 2 major proteins, PFRA and PFRC. We have generated an inducible mutant trypanosome cell line (snl-2) that expresses linked inverted copies of a PFRA gene, capable of forming a PFRA double-stranded (ds) RNA. When expression of this dsRNA was induced, new PFRA RNA and PFRA protein quickly disappeared and PFR construction was affected, resulting in cell paralysis. This inducible RNA interference (RNAi) effect was fast-acting, heritable and reversible. It allowed us to demonstrate that PFR proteins are able to enter both mature and growing flagella but appear to concentrate differentially in new flagella because of the construction process. The PFR is constructed by a polar assembly process at the distal end of the flagellum resulting in a stable cytoskeletal structure with low turn-over. The inducible RNAi approach will have widespread applicability in studies of gene function and cellular processes in parasites. |
| first_indexed | 2024-03-06T18:15:41Z |
| format | Journal article |
| id | oxford-uuid:04884678-e5a5-4b85-8dbe-9d5024ab9ad2 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-06T18:15:41Z |
| publishDate | 2000 |
| record_format | dspace |
| spelling | oxford-uuid:04884678-e5a5-4b85-8dbe-9d5024ab9ad22022-03-26T08:52:17ZFlagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:04884678-e5a5-4b85-8dbe-9d5024ab9ad2EnglishSymplectic Elements at Oxford2000Bastin, PEllis, KKohl, LGull, KThe African trypanosome, Trypanosoma brucei possesses a large and unique intraflagellar structure called the paraflagellar rod (PFR). The PFR is composed of 2 major proteins, PFRA and PFRC. We have generated an inducible mutant trypanosome cell line (snl-2) that expresses linked inverted copies of a PFRA gene, capable of forming a PFRA double-stranded (ds) RNA. When expression of this dsRNA was induced, new PFRA RNA and PFRA protein quickly disappeared and PFR construction was affected, resulting in cell paralysis. This inducible RNA interference (RNAi) effect was fast-acting, heritable and reversible. It allowed us to demonstrate that PFR proteins are able to enter both mature and growing flagella but appear to concentrate differentially in new flagella because of the construction process. The PFR is constructed by a polar assembly process at the distal end of the flagellum resulting in a stable cytoskeletal structure with low turn-over. The inducible RNAi approach will have widespread applicability in studies of gene function and cellular processes in parasites. |
| spellingShingle | Bastin, P Ellis, K Kohl, L Gull, K Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title | Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title_full | Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title_fullStr | Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title_full_unstemmed | Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title_short | Flagellum ontogeny in trypanosomes studied via an inherited and regulated RNA interference system. |
| title_sort | flagellum ontogeny in trypanosomes studied via an inherited and regulated rna interference system |
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