Hatchet ribozyme structure and implications for cleavage mechanism

Zheng, Luqian1; Falschlunger, Christoph2; Huang, Kaiyi1; Mairhofer, Elisabeth2; Yuan, Shuguang3; Wang, Juncheng4; Patel, Dinshaw J.4,5; Micura, Ronald2; Ren, Aiming1

2019-05-28

10.1073/pnas.1902413116

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   影响因子和分区

0027-8424

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-angstrom 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.

hatchet ribozyme cleavage catalysis noncoding RNA

SCI

英语

NI期刊

通讯

NIH[1U19CA179564] ; NIH[P30CA008748]

Science & Technology - Other Topics

Multidisciplinary Sciences

WOS:000469280300034

NATL ACAD SCIENCES

BIOLOGY & BIOCHEMISTRY;CLINICAL MEDICINE;MULTIDISCIPLINARY;PLANT & ANIMAL SCIENCE;ENVIRONMENT/ECOLOGY;SOCIAL SCIENCES, GENERAL;MICROBIOLOGY;ECONOMICS BUSINESS;IMMUNOLOGY;MATERIALS SCIENCE;MATHEMATICS;SPACE SCIENCE;MOLECULAR BIOLOGY & GENETICS;PHARMACOLOGY & TOXICOLOGY;CHEMISTRY;PSYCHIATRY/PSYCHOLOGY;NEUROSCIENCE & BEHAVIOR;PHYSICS;GEOSCIENCES;AGRICULTURAL SCIENCES;ENGINEERING

Web of Science

1.Zhejiang Univ, Life Sci Inst, Hangzhou 310058, Zhejiang, Peoples R China
2.Leopold Franzens Univ, Inst Organ Chem, A-6020 Innsbruck, Austria
3.Ecole Polytech Fed Lausanne, Inst Chem Sci & Engn, CH-1015 Lausanne, Switzerland
4.Mem Sloan Kettering Canc Ctr, Struct Biol Program, New York, NY 10065 USA
5.Southern Univ Sci & Technol, Dept Biol, Shenzhen 518055, Guangdong, Peoples R China

Zheng, Luqian,Falschlunger, Christoph,Huang, Kaiyi,et al. Hatchet ribozyme structure and implications for cleavage mechanism[J]. PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,2019,116(22):10783-10791.

Zheng, Luqian.,Falschlunger, Christoph.,Huang, Kaiyi.,Mairhofer, Elisabeth.,Yuan, Shuguang.,...&Ren, Aiming.(2019).Hatchet ribozyme structure and implications for cleavage mechanism.PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA,116(22),10783-10791.

Zheng, Luqian,et al."Hatchet ribozyme structure and implications for cleavage mechanism".PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA 116.22(2019):10783-10791.