Cryo-EM structures of full-length Tetrahymena ribozyme at 3.1&nbsp;Å resolution

Su, Zhaoming; Zhang, Kaiming; Kappel, Kalli; Li, Shanshan; Palo, Michael Z.; Pintilie, Grigore D.; Rangan, Ramya; Luo, Bingnan; Wei, Yuquan; Das, Rhiju; Chiu, Wah

doi:10.1038/s41586-021-03803-w

Cryo-EM structures of full-length Tetrahymena ribozyme at 3.1 Å resolution

Journal Article · Wed Aug 11 00:00:00 EDT 2021 · Nature (London)

DOI:https://doi.org/10.1038/s41586-021-03803-w· OSTI ID:1818908

^[1]; Zhang, Kaiming ^[2]; ^[3]; ^[2]; Palo, Michael Z. ^[3]; Pintilie, Grigore D. ^[4]; ^[3]; Luo, Bingnan ^[5]; Wei, Yuquan ^[5]; ^[3]; ^[6]

Sichuan Univ., Chengdu (China). West China Hospital, National Clinical Research Center for Geriatrics, State Key Lab. of Biotherapy and Cancer Center; Stanford Univ., CA (United States). James H. Clark Center
Stanford Univ., CA (United States). James H. Clark Center; Univ. of Science and Technology of China, Hefei (China). MOE Key Lab. for Membraneless Organelles and Cellular Dynamics, Hefei National Lab. for Physical Sciences at the Microscale
Stanford Univ., CA (United States)
Stanford Univ., CA (United States). James H. Clark Center
Sichuan Univ., Chengdu (China). West China Hospital, National Clinical Research Center for Geriatrics, State Key Lab. of Biotherapy and Cancer Center
Stanford Univ., CA (United States). James H. Clark Center; SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)

Single-particle cryogenic electron microscopy (cryo-EM) has become a standard technique for determining protein structures at atomic resolution1,2,3. However, cryo-EM studies of protein-free RNA are in their early days. The Tetrahymena thermophila group I self-splicing intron was the first ribozyme to be discovered and has been a prominent model system for the study of RNA catalysis and structure–function relationships4, but its full structure remains unknown. Here we report cryo-EM structures of the full-length Tetrahymena ribozyme in substrate-free and bound states at a resolution of 3.1 Å. Additionally, newly resolved peripheral regions form two coaxially stacked helices; these are interconnected by two kissing loop pseudoknots that wrap around the catalytic core and include two previously unforeseen (to our knowledge) tertiary interactions. The global architecture is nearly identical in both states; only the internal guide sequence and guanosine binding site undergo a large conformational change and a localized shift, respectively, upon binding of RNA substrates. These results provide a long-sought structural view of a paradigmatic RNA enzyme and signal a new era for the cryo-EM-based study of structure–function relationships in ribozymes.

View Accepted Manuscript (DOE)

Research Organization:: SLAC National Accelerator Laboratory (SLAC), Menlo Park, CA (United States)

Sponsoring Organization:: Gabilan Stanford Graduate Fellowship; National Institutes of Health (NIH); National Natural Science Foundation of China (NSFC); National Science Foundation (NSF); USDOE Office of Science (SC), Basic Energy Sciences (BES)

Grant/Contract Number:: AC02-76SF00515

OSTI ID:: 1818908

Alternate ID(s):: OSTI ID: 1839263

Journal Information:: Nature (London), Journal Name: Nature (London) Journal Issue: 7873 Vol. 596; ISSN 0028-0836

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

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Cryo-EM structures of full-length Tetrahymena ribozyme at 3.1 Å resolution

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