vestnik-bio-msuВестник Московского университета. Серия 16. БиологияVestnik Moskovskogo universiteta. Seriya 16. Biologiya0137-0952Lomonosov Moscow State University, School of Biology10.55959/MSU0137-0952-16-80-3S-9vestnik-bio-msu-1542Research ArticleОригинальные исследованияПробоподготовка элонгационного комплекса +39 для криоэлектронной микроскопииSample preparation of the elongation complex +39 for cryo-electron microscopyhttps://orcid.org/0009-0001-0617-9019ОсинаЕ. В.OsinaE. V.

Осина Елизавета Васильевна – мл. науч. сотр. кафедры биоинженерии биологического факультета

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-939-33-59 

 1–73 Leninskie Gory, Moscow, 119234 

osina.elizaveta00@mail.ruhttps://orcid.org/0000-0003-1112-2356МоисеенкоА. В.MoiseenkoA. V.

Моисеенко Андрей Владимирович – науч. сотр. кафедры биоинженерии биологического факультета 

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-939-33-59 

 1–73 Leninskie Gory, Moscow, 119234 

postmoiseenko@gmail.comhttps://orcid.org/0000-0001-9240-9054КоровинаА. Н.KorovinaA. N.

Коровина Анна Николаевна – канд. биол. наук, мл. науч. сотр. кафедры биоинженерии биологического факультета

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-939-57-38  

 1–73 Leninskie Gory, Moscow, 119234 

anna.korovina@gmail.comhttps://orcid.org/0000-0002-5048-0378ГерасимоваН. С.GerasimovaN. S.

Герасимова Надежда Сергеевна – канд. биол. наук, ст. науч. сотр. кафедры биоинженерии биологического факультета 

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-938-22-91 

 1–73 Leninskie Gory, Moscow, 119234 

shordome@gmail.comhttps://orcid.org/0000-0003-2261-9587ВолохО. И.VolokhO. I.

Волох Олеся Игоревна – канд. биол. наук, науч. сотр. кафедры биоинженерии биологического факультета 

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-939-57-38 

 1–73 Leninskie Gory, Moscow, 119234 

olesyavolokh@gmail.comhttps://orcid.org/0000-0002-7389-7993СтудитскийВ. М.StuditskyV. M.

Студитский Василий Михайлович – докт. биол. наук, вед. науч. сотр. кафедры биоинженерии биологического факультета 

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

333 Cottman Ave., Philadelphia, 19111, Pennsylvania 

Тел.: 8-495-939-57-38 

 1–73 Leninskie Gory, Moscow, 119234 

 333 Cottman Ave., Philadelphia, 19111, Pennsylvania 

vasily.studitsky@fccc.eduhttps://orcid.org/0000-0002-8427-8178Станишнева-КоноваловаТ. Б.Stanishneva-KonovalovaT. B.

Станишнева-Коновалова Татьяна Борисовна – канд. биол. наук, ст. науч. сотр. кафедры биоинженерии биологического факультета 

119234, г. Москва, Ленинские горы, д. 1, стр. 73 

Тел.: 8-495-939-57-38 

 1–73 Leninskie Gory, Moscow, 119234 

stanishneva-konovalova@mail.bio.msu.ruКафедра биоинженерии, биологический факультет, Московский государственный университет имени М.В. ЛомоносоваРоссияDepartment of Bioengineering, Faculty of Biology, Lomonosov Moscow State UniversityRussian FederationКафедра биоинженерии, биологический факультет, Московский государственный университет имени М.В. Ломоносова; Fox Chase Cancer CenterРоссияDepartment of Bioengineering, Faculty of Biology, Lomonosov Moscow State University; Fox Chase Cancer CenterRussian Federation202510112025803S59–66Copyright © Осина Е.В., Моисеенко А.В., Коровина А.Н., Герасимова Н.С., Волох О.И., Студитский В.М., Станишнева-Коновалова Т.Б., 20252025Осина Е.В., Моисеенко А.В., Коровина А.Н., Герасимова Н.С., Волох О.И., Студитский В.М., Станишнева-Коновалова Т.Б.Osina E.V., Moiseenko A.V., Korovina A.N., Gerasimova N.S., Volokh O.I., Studitsky V.M., Stanishneva-Konovalova T.B.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestnik-bio-msu.elpub.ru/jour/article/view/1542

Транскрипция в клетке осуществляется специализированными ферментами – РНК-полимеразами. РНК-полимеразы транскрибируют ДНК с образованием элонгационных комплексов (ЭК), в том числе участвующих в регуляции транскрипции. Криоэлектронная микроскопия (криоЭМ) позволяет определять структуры этих комплексов и выяснять механизмы транскрипции хроматина. Однако подготовка образцов ЭК, пригодных для изучения методом криоЭМ, представляет определенные сложности. В данной работе проведена разработка протоколов для подготовки образов ЭК с положением активного центра РНКП в позиции +39 от входа в нуклеосому (ЭК+39). Образование комплексов подтверждалось методами электрофореза и электронной микроскопии негативного контрастирования. Разработанные методы могут быть использованы для изучения ЭК+39 методом криоЭМ.

Transcription in the cell is carried out by specialized enzymes – RNA polymerases. RNA polymerases transcribe DNA with the formation of elongation complexes (EC), which have a regulatory significance. Cryo-electron microscopy (cryo-EM) allows obtaining the structures of these complexes and clarifying the mechanisms of transcription stages. However, the preparation of EC samples suitable for cryo-EM studies presents certain challenges. In this work, a selection of protocols for the preparation of EC+39 samples was carried out. The formation of complexes is confirmed by the results of electrophoresis and negative stain electron microscopy. The results obtained can be used to study EC+39 by the cryo-EM method.

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