Исследование бинарных комплексов РНК и рибосомного белка S7 эубактерий

6 ВЫВОДЫ:

1. Методами компьютерного анализа и моделирования in silico показано, что два белка-аналога EcoS7 и TthS7 по первичной структуре идентичны на 52%, а в зоне РНК-белковых контактов в 30S субчастице — на 83%. Подобная высокая консервативность дает возможность экспериментально сравнить свойства гомологичных и гетерологичных РНК-белковых комплексов, причем TthS7 можно рассматривать как «природный мутант» EcoS7.

2. Получен супер-продуцент Е. coli для EcoS7. Показано, что для стабильной экспрессии регуляторного белка необходимы системы с самым жестким контролем регуляции экспрессии (pET28b/BL21 DE3 pLysE).

3. Установлено, что взаимодействие in vitro EcoS7 и TthS7 с фрагментом 16S рРНК, в целом однотипно, но отлично от таковых в 30S субчастице рибосом.

4. Установлено, что белок S7 взаимодействует с участком бифуркации межцистронного фрагмента S12-S7 str мРНК, что приводит к изменению конформации в основании шпильки, где расположены сигналы трансляции. На основании этого предложена гипотеза о механизме регуляции сопряженной трансляции.

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