Biopolym. Cell. 2002; 18(2):142-154.
Роль вышерасположенного стимулирующего фактора-2а в регуляции экспрессии ингибитора активатора плазминогена-1
1Самойленко А. А., 1Рот У., 1Юнгерманн К., 1Китцманн Т., 2Оболенская М. Ю.
  1. Институт биохимии и молекулярной биологии клетки
    Гумбольдталее 23, D-37073 Геттинген, Германия
  2. Институт молекулярной биологии и генетики НАН Украины
    ул. Академика Заболотного, 150, Киев, Украина, 03680

Abstract

Экспрессия ингибитора активатора плазминогена-1 (ИАП-1) стимулируется гипоксией (8 % О2) За эту стимуляцию отвечает участок в промоторе ИАП-1 (-175/-159), содержа­щий элемент, воспринимающий гипоксию (ЭВГ-2). С послед­ним взаимодействует фактор 1, стимулируемый гипоксией (ФСГ-1). С участком ЭВГ-2 граничит область ЭВГ-1, с которой связывается ранее не идентифицированный фактор. В данной работе осуществлена попытка идентифицировать этот фактор и выяснить его роль в регуляции экспрессии ИАП-1. На основании сдвига специфической электрофоретической полосы доказано, что с сайтами ЭВГ, преимущественно с ЭВГ-1, связывается вышерасположенный стимулирующий фактор-2а, ВСФ-2а. В первичной культуре гепатоцитов кры­сы суперэкспрессия ВСФ-2а в одинаковой мере угнетала экспрессию ИАП-1-специфической мРНК и белка ИАП-1 в услови­ях как гипоксии, так и нормоксии. Первичные гепатоциты крысы трансфицировали двумя рекомбинантными ДНК – кон­струкцией с геном люциферази под контролем участка промо­тора ИАП-1 длиной 766 или 276 п. о. и векторами, экспрессирующими ВСФ дикого (ВСФ-2а) или мутантного (ΔВСФ-2а) типа. Последний не выявляет ДНК-связывающей и димеризационной активностей. Котрансфекция с вектором, экспрессирующим ВСФ-2а, угнетала активность люциферазы в 8 раз по отношению к контролю, где белок ВСФ-2а отсутствовал Котрансфекция с вектором, экспрессирующим ΔВСФ-2а, не влияла на люциферазную активность. Мутации обоих ЭВГ и в большей мере мутация ЭВГ-1 препятствовали угнетению контролированной промотором ИАП-1 люциферазной актив­ности. Ингибитор ного действия ВСФ-2а, опосредованного ЭВГ-2, в отличие от ЭВГ-1, не наблюдалось при суперэкспрес­сии ФСГ-la. Эти результаты свидетельствуют о том, что баланс между двумя транскрипционными факторами, ВСФ-2а и ФСГ-la, способными связываться со смежными ЭВГ, играет определенную роль в регуляции экспрессии ИАП-1 в условиях гипо- и нормоксии.

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