БИОХИМИЯ, 2022, том 87, вып. 1, с. 3–15

УДК 577.123

Влияние метилирования ДНК на 3′→5′-экзонуклеазную активность основной апурин-апиримидиновой эндонуклеазы человека APEX1

© 2022 А.В. Ендуткин 1, Д.Д. Яценко 1, Д.О. Жарков 1,2*dzharkov@niboch.nsc.ru

1 Институт химической биологии и фундаментальной медицины СО РАН, 630090 Новосибирск, Россия

2 Новосибирский государственный университет, факультет естественных наук, 630090 Новосибирск, Россия

Поступила в редакцию 28.07.2021
После доработки 23.11.2021
Принята к публикации 24.11.2021

DOI: 10.31857/S0320972522010018

КЛЮЧЕВЫЕ СЛОВА: повреждение ДНК, репарация ДНК, эпигенетическое деметилирование, АП‑эндонуклеаза, 3′→5′-экзонуклеаза, APEX1, 5‑метилцитозин.

Аннотация

Апурин-апиримидиновые (АП) эндонуклеазы – ключевые ферменты пути эксцизионной репарации оснований ДНК, гидролизующие фосфодиэфирную связь АП‑сайта, который образуется после удаления повреждённого основания. Основная АП‑эндонуклеаза человека APEX1, помимо этого, проявляет 3′‑фосфодиэстеразную и 3′→5′-экзонуклеазную активности. Биологические функции последней не выяснены; предполагается, что она может корректировать ошибки репаративного синтеза ДНК. При повреждении ДНК с 3′‑стороны вблизи 5‑метилцитозина (mC) проявление 3′→5′-экзонуклеазной активности может вызывать изменение статуса эпигенетического метилирования CpG-динуклеотидов. Остаётся открытым вопрос: вносит ли 3′→5′-экзонуклеазная активность APEX1 вклад в процесс активного эпигенетического деметилирования или, напротив, она лимитирована в метилированных CpG-динуклеотидах для сохранения эпигенетического статуса при репарации случайных повреждений ДНК. В работе впервые систематически исследована эффективность удаления ферментом APEX1 3′‑концевых нуклеотидов из субстратов, представляющих собой интермедиаты репарации ДНК в CpG-динуклеотидах. Показано, что во всех интермедиатах олигонуклеотиды с некомплементарными матрице основаниями на 3′ конце представляют собой наилучшие, а с mC – наихудший субстрат для проявления 3′→5′-экзонуклеазной активности APEX1. Наличие mC в комплементарной цепи значительно снижает константу скорости реакции даже для некомплементарных 3′‑концевых звеньев. Таким образом, эффективность 3′→5′-экзонуклеазной реакции, катализируемой APEX1, ограничена в метилированных CpG-динуклеотидах, что, вероятно, связано с необходимостью поддержания эпигенетического статуса при репарации.

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Финансирование

Исследование поддержано Российским научным фондом (грант № 17‑14‑01190П, экспериментальная биохимическая часть) и государственным заданием 0245‑2021‑0002 (структурный анализ).

Конфликт интересов

Авторы заявляют об отсутствии конфликта интересов.

Соблюдение этических норм

Настоящая статья не содержит описания каких-либо исследований с участием людей или животных в качестве объектов.

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