Изучение взаимодействий РНК-полимеразы с однонитевыми и двунитевыми ДНК-аптамерами
Michaud M., Jourdan E., Ravelet C., Villet A., Ravel A., Grosset C., Peyrin E. 2004. Immobilized DNA aptamers as target-specific chiral stationary phases for resolution of nucleoside and amino acid derivative enantiomers. Anal. Chem. 76: 1015−20. Convery, M.A., Rowsell, S., Stonehouse, N.J., Ellington, A.D., Hirao, I., Murray, J.B., Peabody, D.S., Phillips, S.E., Stockley, P.G. 1998. Crystal… Читать ещё >
Изучение взаимодействий РНК-полимеразы с однонитевыми и двунитевыми ДНК-аптамерами (реферат, курсовая, диплом, контрольная)
Содержание
- Выводы
1. Получены и охарактеризованы высоаффинные ДНК-аптамеры к кор-РНК-полимеразам Е. coli и Т. aquaticus, которые являются эффективными ингибиторами транскрипции. При помощи аптамеров показано, что фактор GreB вызывает конформационные перестройки внутри главного канала РНКП.
2. Осуществлён поиск ДНК-аптамеров к изолированной а-субъединице РНК-полимеразы Т. aquaticus. Отобранные аптамеры содержат последовательность нематричной нити -10 области бактериальных промоторов. Таким образом, свободная а-субъединица способна узнавать промоторную последовательность в отсутствие кор-фермента РНК-полимеразы. Аптамеры взаимодействуют с РНК-полимеразой с высокой аффинностью и являются эффективными ингибиторами синтеза РНК in vitro.
3. Двунитевой фрагмент ДНК, содержащий последовательность аптамера к ст-субъединице Т. aquaticus, является промотором нового типа, не содержащим -35 области и TG-элемента и специфичным для холофермента РНК-полимеразы Т. aquaticus. Для узнавания аптамерного промотора необходимо наличие расширенного мотива -10 области TATAATGGGA, содержащего ранее не известный GGGA-элемент.
4. Однонитевой ДНК-аптамер к изолированной <�т-субъединице Т. aquaticus служит матрицей для специфической ст-зависимой инициации транскрипции РНК-полимеразой Т. aquaticus.
Список работ, опубликованных по теме диссертации
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Feklistov A., Kulbachinsky A., Nikiforov V. DNA aptamers unveil sequence specificity of free sigma factor. FASEB Summer Research Conference «Transcription initiation in Prokaryotes». June 12−17, 2004, Vermont Academy in Saxtons River, Vermont, USA.
Феклистов А. В., Кульбачинский А. В., Никифоров В. Г. ДПК-связывающая специфичность ст-субъединицы РНК-полимеразы Т. aquaticus, изученная методом SELEX. Конференция молодых учёных, аспирантов и студентов по молекулярной биологии и генетике, посвященная 50-летию открытия двойной спирали ДНК, а также 30-ой годовщине Института молекулярной биологии и генетики академии наук Украины. 25−27 сентября 2003 г., Украина, Киев.
Благодарности
Автор благодарит А. В. Кульбачинского и В. Г. Никифорова за организацию работы, руководство, советы и критику, А. Гольдфарба и И. А. Крашенинникова за создание условий для работы.
Автор благодарен сотрудникам ЛМГМ, ОМГЖ ИМГ РАН и лаборатории молекулярной биофизики Рокфеллеровского университета, а также коллективу Кафедры молекулярной биологии Биологического факультета МГУ за внимание, советы и помощь в работе.
Автор очень признателен Н. Наумовой, С. Колбу, А. Кульбачинскому, Н. Севостьяновой, В. Вагину, А. и П. Моториным, М. и М. Фрих-Харам, А. Мустаеву, И. Аджубею, И. Абрамсон, А. Машину, О. Глебову, А. Ленцу, В. Эпштейну, Д. Альперну, Л. Вестблэйд и Д. Мюррей за неоценимую дружескую помощь и поддержку в процессе выполнения работы.
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