vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-23-112vavilov-4008Research ArticleГЕНЕТИКА РАЗВИТИЯGENETIC ENGINEERINGИнактивация компонентов комплекса TIM приводит к снижению уровня импорта ДНК в митохондрии арабидопсисаInactivation of the TIM complex components leads to a decrease in the level of DNA import into Arabidopsis mitochondriahttps://orcid.org/0000-0002-2830-4175ТарасенкоТ. А.TarasenkoT. A.

Иркутск

Irkutsk

ЕлизоваК. Д.ElizovaK. D.

Иркутск

Irkutsk

https://orcid.org/0000-0001-8208-6941ТарасенкоВ. И.TarasenkoV. I.

Иркутск

Irkutsk

vslav@inbox.ruhttps://orcid.org/0000-0003-0931-2863КулинченкоМ. В.KoulintchenkoM. V.

Иркутск

Казань

Irkutsk

Kazan

https://orcid.org/0000-0002-0601-2788КонстантиновЮ. М.KonstantinovYu. M.

Иркутск

Irkutsk

Сибирский институт физиологии и биохимии растений Сибирского отделения Российской академии наукРоссияSiberian Institute of Plant Physiology and Biochemistry of the Siberian Branch of the Russian Academy of SciencesRussian FederationСибирский институт физиологии и биохимии растений Сибирского отделения Российской академии наук; Казанский институт биохимии и биофизики Казанского научного центра Российской академии наукРоссияSiberian Institute of Plant Physiology and Biochemistry of the Siberian Branch of the Russian Academy of Sciences; Kazan Institute of Biochemistry and Biophysics of Kazan Scientific Center of the Russian Academy of SciencesRussian Federation202329122023278971979Copyright © Тарасенко Т.А., Елизова К.Д., Тарасенко В.И., Кулинченко М.В., Константинов Ю.М., 20232023Тарасенко Т.А., Елизова К.Д., Тарасенко В.И., Кулинченко М.В., Константинов Ю.М.Tarasenko T.A., Elizova K.D., Tarasenko V.I., Koulintchenko M.V., Konstantinov Y.M.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/4008

Феномен импорта ДНК в митохондрии показан для всех основных групп эукариот. В растениях и животных импорт ДНК, по-видимому, происходит различными путями. Известно, что в растительные органеллы нуклеиновые кислоты попадают по альтернативным каналам в зависимости от размера импортируемых молекул. Импорт ДНК небольшого размера (до 300 п. н.) частично перекрывается с механизмом импорта тРНК, по крайней мере, на уровне внешней мембраны. Примечательно, что у растений в импорт тРНК вовлечены компоненты аппарата импорта белков, чья роль в транспорте ДНК до настоящего времени оставалась неизученной. В настоящей работе мы провели исследование роли отдельных компонентов транслоказы внутренней мембраны TIM в процессе импорта ДНК в изолированные митохондрии арабидопсиса и их возможной связи с порином VDAC1. С использованием нокаут-мутантов по генам, кодирующим изоформы белков Tim17 или Tim23, мы впервые показали участие этих белков в импорте фрагментов ДНК разной длины. Кроме того, ингибирование транспортных каналов специфическими антителами к VDAC1 приводило к снижению уровня импорта ДНК в митохондрии дикого типа, что позволило установить специфическое участие этой изоформы порина в импорте ДНК. В нокаут-мутанте tim17-1 происходило дополнительное снижение эффективности импорта ДНК в присутствии антител к VDAC1 в сравнении с линией дикого типа. Полученные результаты указывают на участие белков Tim17-1 и Tim23-2 в аппарате импорта ДНК в растительные митохондрии. При этом Tim23-2 может быть часть канала, формируемого при участии VDAC1, в то время как Tim17-1, по-видимому, вовлечен в альтернативный, независимый от VDAC1, путь импорта ДНК. Выявление мембранных белков-переносчиков, участвующих в различных путях импорта ДНК, позволит использовать природную способность митохондрий к поглощению ДНК в качестве удобного биотехнологического инструмента для трансформации митохондриального генома.

The phenomenon of DNA import into mitochondria has been shown for all major groups of eukaryotes. In plants and animals, DNA import seems to occur in different ways. It has been known that nucleic acids enter plant organelles through alternative channels, depending on the size of the imported molecules. Mitochondrial import of small DNA (up to 300 bp) partially overlaps with the mechanism of tRNA import, at least at the level of the outer membrane. It is noteworthy that, in plants, tRNA import involves components of the protein import apparatus, whose role in DNA transport has not yet been studied. In this work, we studied the role of individual components of the TIM inner membrane translocase in the process of DNA import into isolated Arabidopsis mitochondria and their possible association with the porin VDAC1. Using knockout mutants for the genes encoding Tim17 or Tim23 protein isoforms, we demonstrated for the first time the involvement of these proteins in the import of DNA fragments of different lengths. In addition, inhibition of transport channels with specific antibodies to VDAC1 led to a decrease in the level of DNA import into wild-type mitochondria, which made it possible to establish the specific involvement of this porin isoform in DNA import. In the tim17-1 knockout mutant, there was an additional decrease in the efficiency of DNA import in the presence of antibodies to VDAC1 compared to the wild type line. The results obtained indicate the involvement of the Tim17-1 and Tim23-2 proteins in the mechanism of DNA import into plant mitochondria. At the same time, Tim23-2 may be part of the channel formed with the participation of VDAC1, while Tim17-1, apparently, is involved in an alternative DNA import pathway independent of VDAC1. The identification of membrane carrier proteins involved in various DNA import pathways will make it possible to use the natural ability of mitochondria to import DNA as a convenient biotechnological tool for transforming the mitochondrial genome.

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The authors declare that there are no conflicts of interest present.