SHAIKH, Tanvir, A.S. YASSIN, Z.H. LU, D. BARNARD, X. MENG, T.M. LU, T. WAGENKNECHT and R. K. AGRAWAL. Initial bridges between two ribosomal subunits are formed within 9.4 milliseconds, as studied by time-resolved cryo-EM. Proceedings of the National Academy of Sciences of the United States of America. WASHINGTON: NATL ACAD SCIENCES, 2014, vol. 111, No 27, p. 9822-9827. ISSN 0027-8424. doi:10.1073/pnas.1406744111.
Other formats:   BibTeX LaTeX RIS
Basic information
Original name Initial bridges between two ribosomal subunits are formed within 9.4 milliseconds, as studied by time-resolved cryo-EM
Authors SHAIKH, Tanvir (840 United States of America, guarantor, belonging to the institution), A.S. YASSIN (840 United States of America), Z.H. LU (840 United States of America), D. BARNARD (840 United States of America), X. MENG (840 United States of America), T.M. LU (840 United States of America), T. WAGENKNECHT (840 United States of America) and R. K. AGRAWAL (840 United States of America).
Edition Proceedings of the National Academy of Sciences of the United States of America, WASHINGTON, NATL ACAD SCIENCES, 2014, 0027-8424.
Other information
Original language English
Type of outcome Article in a journal
Field of Study 10600 1.6 Biological sciences
Country of publisher United States of America
Confidentiality degree is not subject to a state or trade secret
WWW URL
Impact factor Impact factor: 9.674
RIV identification code RIV/00216224:14740/14:00079214
Organization unit Central European Institute of Technology
Doi http://dx.doi.org/10.1073/pnas.1406744111
UT WoS 000338514800036
Keywords in English ribosomal intersubunit bridges; millisecond time resolution cryo-EM
Tags kontrola MP, MP, OA, rivok, SCOPUS
Tags International impact, Reviewed
Changed by Changed by: Martina Prášilová, učo 342282. Changed: 25. 2. 2015 09:03.
Abstract
Association of the two ribosomal subunits during the process of translation initiation is a crucial step of protein synthesis. The two subunits (30S and 50S) of the bacterial 70S ribosome are held together by 12 dynamic bridges involving RNA-RNA, RNA-protein, and protein-protein interactions. The process of bridge formation, such as whether all these bridges are formed simultaneously or in a sequential order, is poorly understood. To understand such processes, we have developed and implemented a class of microfluidic devices that mix two components to completion within 0.4 ms and spray the mixture in the form of microdroplets onto an electron microscopy grid, yielding a minimum reaction time of 9.4 ms before cryofixation. Using these devices, we have obtained cryo-EM data corresponding to reaction times of 9.4 and 43 ms and have determined 3D structures of ribosomal subunit association intermediates. Molecular analyses of the cryo-EM maps reveal that eight intersubunit bridges (bridges B1a, B1b, B2a, B2b, B3, B7a, B7b, and B8) form within 9.4 ms, whereas the remaining four bridges (bridges B2c, B4, B5, and B6) take longer than 43 ms to form, suggesting that bridges are formed in a stepwise fashion. Our approach can be used to characterize sequences of various dynamic functional events on complex macromolecular assemblies such as ribosomes.
Links
EE2.3.20.0042, research and development projectName: Internacionalizace programu Strukturní biologie s důrazem na rozvoj nových směrů výzkumu
PrintDisplayed: 21. 5. 2022 22:09