Dynamic topology of double-stranded telomeric DNA studied by single-molecule manipulation in vitro.

Abstract:

:The dynamic topological structure of telomeric DNA is closely related to its biological function; however, no such structural information on full-length telomeric DNA has been reported due to difficulties synthesizing long double-stranded telomeric DNA. Herein, we developed an EM-PCR and TA cloning-based approach to synthesize long-chain double-stranded tandem repeats of telomeric DNA. Using mechanical manipulation assays based on single-molecule atomic force microscopy, we found that mechanical force can trigger the melting of double-stranded telomeric DNA and the formation of higher-order structures (G-quadruplexes or i-motifs). Our results show that only when both the G-strand and C-strand of double-stranded telomeric DNA form higher-order structures (G-quadruplexes or i-motifs) at the same time (e.g. in the presence of 100 mM KCl under pH 4.7), that the higher-order structure(s) can remain after the external force is removed. The presence of monovalent K+, single-wall carbon nanotubes (SWCNTs), acidic conditions, or short G-rich fragments (∼30 nt) can shift the transition from dsDNA to higher-order structures. Our results provide a new way to regulate the topology of telomeric DNA.

journal_name

Nucleic Acids Res

journal_title

Nucleic acids research

authors

Zhang X,Zhang Y,Zhang W

doi

10.1093/nar/gkaa479

subject

Has Abstract

pub_date

2020-07-09 00:00:00

pages

6458-6470

issue

12

eissn

0305-1048

issn

1362-4962

pii

5851393

journal_volume

48

pub_type

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