Dynamic structural insights into the molecular mechanism of DNA unwinding by the bacteriophage T7 helicase.

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 Res

journal_title

Nucleic acids research

authors

Ma JB,Chen Z,Xu CH,Huang XY,Jia Q,Zou ZY,Mi CY,Ma DF,Lu Y,Zhang HD,Li M

doi

10.1093/nar/gkaa057

subject

Has Abstract

pub_date

2020-04-06 00:00:00

pages

3156-3164

issue

6

eissn

0305-1048

issn

1362-4962

pii

5721210

journal_volume

48

pub_type

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