Abstract:
:The hexametric T7 helicase (gp4) adopts a spiral lock-washer form and encircles a coil-like DNA (tracking) strand with two nucleotides bound to each subunit. However, the chemo-mechanical coupling mechanism in unwinding has yet to be elucidated. Here, we utilized nanotensioner-enhanced Förster resonance energy transfer with one nucleotide precision to investigate gp4-induced unwinding of DNA that contains an abasic lesion. We observed that the DNA unwinding activity of gp4 is hindered but not completely blocked by abasic lesions. Gp4 moves back and forth repeatedly when it encounters an abasic lesion, whereas it steps back only occasionally when it unwinds normal DNA. We further observed that gp4 translocates on the tracking strand in step sizes of one to four nucleotides. We propose that a hypothetical intermediate conformation of the gp4-DNA complex during DNA unwinding can help explain how gp4 molecules pass lesions, providing insights into the unwinding dynamics of gp4.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Ma JB,Chen Z,Xu CH,Huang XY,Jia Q,Zou ZY,Mi CY,Ma DF,Lu Y,Zhang HD,Li Mdoi
10.1093/nar/gkaa057subject
Has Abstractpub_date
2020-04-06 00:00:00pages
3156-3164issue
6eissn
0305-1048issn
1362-4962pii
5721210journal_volume
48pub_type
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