| Author |
Rao, Zihe Sun, Yuna Ma, Yuanyuan Shaw, Neil Gao, Yan Wang, Jin Lou, Zhiyong Zhang, Rongguang Wu, Lijie Yan, Liming |
| Description |
Author Affiliation: Ma Y ( Laboratory of Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China); Wu L ( National Center for Protein Science Shanghai, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China); Shaw N ( National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China); Gao Y ( Laboratory of Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China); Wang J ( State Key Laboratory of Biotherapy and Cancer Center, Sichuan University, Chengdu, Sichuan 610041, China); Sun Y ( National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China); Lou Z ( Laboratory of Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China); Yan L ( Laboratory of Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China); Zhang R ( National Center for Protein Science Shanghai, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China); Rao Z ( Laboratory of Structural Biology, School of Life Sciences, Tsinghua University, Beijing 100084, China); |
| Abstract |
Nonstructural protein 14 (nsp14) of coronaviruses (CoV) is important for viral replication and transcription. The N-terminal exoribonuclease (ExoN) domain plays a proofreading role for prevention of lethal mutagenesis, and the C-terminal domain functions as a (guanine-N7) methyl transferase (N7-MTase) for mRNA capping. The molecular basis of both these functions is unknown. Here, we describe crystal structures of severe acute respiratory syndrome (SARS)-CoV nsp14 in complex with its activator nonstructural protein10 (nsp10) and functional ligands. One molecule of nsp10 interacts with ExoN of nsp14 to stabilize it and stimulate its activity. Although the catalytic core of nsp14 ExoN is reminiscent of proofreading exonucleases, the presence of two zinc fingers sets it apart from homologs. Mutagenesis studies indicate that both these zinc fingers are essential for the function of nsp14. We show that a DEEDh (the five catalytic amino acids) motif drives nucleotide excision. The N7-MTase domain exhibits a noncanonical MTase fold with a rare ß-sheet insertion and a peripheral zinc finger. The cap-precursor guanosine-P3-adenosine-5',5'-triphosphate and S-adenosyl methionine bind in proximity in a highly constricted pocket between two ß-sheets to accomplish methyl transfer. Our studies provide the first glimpses, to our knowledge, into the architecture of the nsp14-nsp10 complex involved in RNA viral proofreading. |
| Subject Keyword |
Exoribonucleases Chemistry SARS Virus Viral Nonstructural Proteins Amino Acid Sequence Escherichia Coli Metabolism Ligands Methyltransferases Models, Molecular Molecular Sequence Data Protein Structure, Secondary Protein Structure, Tertiary RNA, Messenger Genetics RNA, Viral Sequence Homology, Amino Acid Virus Replication Zinc Fingers Research Support, Non-U.S. Gov't Multidisciplinary |
