Sample preparation of the elongation complex +39 for cryo-electron microscopy
E. V. Osina,
A. V. Moiseenko,
A. N. Korovina,
N. S. Gerasimova,
O. I. Volokh,
V. M. Studitsky,
T. B. Stanishneva-Konovalova https://doi.org/10.55959/MSU0137-0952-16-80-3S-9
Abstract
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.
About the Authors
E. V. Osina
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
A. V. Moiseenko
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
A. N. Korovina
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
N. S. Gerasimova
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
O. I. Volokh
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
V. M. Studitsky
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University; Fox Chase Cancer Center
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
333 Cottman Ave., Philadelphia, 19111, Pennsylvania
T. B. Stanishneva-Konovalova
Department of Bioengineering, Faculty of Biology, Lomonosov Moscow State University
Russian Federation
Russian Federation
1–73 Leninskie Gory, Moscow, 119234
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Review
For citations:
Osina E.V., Moiseenko A.V., Korovina A.N., Gerasimova N.S., Volokh O.I., Studitsky V.M., Stanishneva-Konovalova T.B. Sample preparation of the elongation complex +39 for cryo-electron microscopy. Vestnik Moskovskogo universiteta. Seriya 16. Biologiya. 2025;80(3S):59–66. (In Russ.) https://doi.org/10.55959/MSU0137-0952-16-80-3S-9
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