Duplex Unwinding Mechanism of Coronavirus MERS-CoV nsp13 Helicase.

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Tác giả: Hamza Balci, Qixin Chen, Sheng Cui, Jiajie Diao, Wei Hao, Pengjiao Hou, Xiao Hu, Ziheng Li, Bo Qin, Zhiqi Tian, Rong Zhao

Ngôn ngữ: eng

Ký hiệu phân loại:

Thông tin xuất bản: United States : Chemical & biomedical imaging , 2025

Mô tả vật lý:

Bộ sưu tập: NCBI

ID: 728156

The COVID-19 pandemic has underscored the importance of in-depth research into the proteins encoded by coronaviruses (CoV), particularly the highly conserved nonstructural CoV proteins (nsp). Among these, the nsp13 helicase of severe pathogenic MERS-CoV, SARS-CoV-2, and SARS-CoV is one of the most preserved CoV nsp. Utilizing single-molecule FRET, we discovered that MERS-CoV nsp13 unwinds DNA in distinct steps of about 9 bp when ATP is employed. If a different nucleotide is introduced, these steps diminish to 3-4 bp. Dwell-time analysis revealed 3-4 concealed steps within each unwinding process, which suggests the hydrolysis of 3-4 dTTP. Combining our observations with previous studies, we propose an unwinding model of CoV nsp13 helicase. This model suggests that the elongated and adaptable 1B-stalk of nsp13 may enable the 1B remnants to engage with the unwound single-stranded DNA, even as the helicase core domain has advanced over 3-4 bp, thereby inducing accumulated strain on the nsp13-DNA complex. Our findings provide a foundational framework for determining the unwinding mechanism of this unique helicase family.
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