Hatchet ribozyme structure and implications for cleavage mechanism

Zheng, Luqian; Falschlunger, Christoph; Huang, Kaiyi; Mairhofer, Elisabeth; Yuan, Shuguang; Wang, Juncheng; Patel, Dinshaw J.; Micura, Ronald; Ren, Aiming

doi:10.1073/pnas.1902413116

Hatchet ribozyme structure and implications for cleavage mechanism

Journal Article · Tue May 14 00:00:00 EDT 2019 · Proceedings of the National Academy of Sciences of the United States of America

DOI:https://doi.org/10.1073/pnas.1902413116· OSTI ID:1531009

Zheng, Luqian ^[1]; Falschlunger, Christoph ^[2]; Huang, Kaiyi ^[1]; Mairhofer, Elisabeth ^[2]; ^[3]; Wang, Juncheng ^[4]; Patel, Dinshaw J. ^[5]; Micura, Ronald ^[2]; ^[1]

Zhejiang Univ., Hangzhou (China). Life Science Inst.
Leopold Franzens Univ., Innsbruck (Austria). Inst. of Organic Chemistry
Ecole Polytechnique Federale Lausanne (Switzlerland)
Memorial Sloan-Kettering Cancer Center, New York, NY (United States)
Memorial Sloan-Kettering Cancer Center, New York, NY (United States); Southern Univ. of Science and Technology, Shenzhen (China)

Small self-cleaving ribozymes catalyze site-specific cleavage of their own phosphodiester backbone with implications for viral genome replication, pre-mRNA processing, and alternative splicing. We report on the 2.1-Å crystal structure of the hatchet ribozyme product, which adopts a compact pseudosymmetric dimeric scaffold, with each monomer stabilized by long-range interactions involving highly conserved nucleotides brought into close proximity of the scissile phosphate. Strikingly, the catalytic pocket contains a cavity capable of accommodating both the modeled scissile phosphate and its flanking 5′ nucleoside. The resulting modeled precatalytic conformation incorporates a splayed-apart alignment at the scissile phosphate, thereby providing structure-based insights into the in-line cleavage mechanism. We identify a guanine lining the catalytic pocket positioned to contribute to cleavage chemistry. The functional relevance of structure-based insights into hatchet ribozyme catalysis is strongly supported by cleavage assays monitoring the impact of selected nucleobase and atom-specific mutations on ribozyme activity.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: National Institutes of Health (NIH); USDOE

OSTI ID:: 1531009

Journal Information:: Proceedings of the National Academy of Sciences of the United States of America, Journal Name: Proceedings of the National Academy of Sciences of the United States of America Journal Issue: 22 Vol. 116; ISSN 0027-8424

Publisher:: National Academy of SciencesCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
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noncoding RNA
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Hatchet ribozyme structure and implications for cleavage mechanism

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