Publication:
A mechanism for the extension and unfolding of parallel telomeric g-quadruplexes by human telomerase at single-molecule resolution

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Issued Date

2020

Resource Type

Article

File Type

application/pdf

ISSN

2050084X

DOI

10.7554/eLife.56428

Other identifier(s)

2-s2.0-85089610831

Rights Holder(s)

Scopus

Bibliographic Citation

eLife. Vol 9, (2020), p.1-9

Suggested Citation

Paudel B.P., Moye A.L., Assi H.A., El-Khoury R., Cohen S.B., Holien J.K., Birrento M.L., Samosorn S., Intharapichai K., Tomlinson C.G., Teulade-Fichou M.-P., González C., Beck J.L., Damha M.J., Van Oijen A.M., Bryan T.M. A mechanism for the extension and unfolding of parallel telomeric g-quadruplexes by human telomerase at single-molecule resolution. eLife. Vol 9, (2020), p.1-9. doi:10.7554/eLife.56428 Retrieved from: https://hdl.handle.net/20.500.14740/4517

Title

A mechanism for the extension and unfolding of parallel telomeric g-quadruplexes by human telomerase at single-molecule resolution

Author(s)

Paudel B.P.
 Moye A.L.
 Assi H.A.
 El-Khoury R.
 Cohen S.B.
 Holien J.K.
 Birrento M.L.
 Samosorn S.
 Intharapichai K.
 Tomlinson C.G.
 Teulade-Fichou M.-P.
 González C.
 Beck J.L.
 Damha M.J.
 Van Oijen A.M.
 Bryan T.M.

Abstract

Telomeric G-quadruplexes (G4) were long believed to form a protective structure at telomeres, preventing their extension by the ribonucleoprotein telomerase. Contrary to this belief, we have previously demonstrated that parallel-stranded conformations of telomeric G4 can be extended by human and ciliate telomerase. However, a mechanistic understanding of the interaction of telomerase with structured DNA remained elusive. Here, we use single-molecule fluorescence resonance energy transfer (smFRET) microscopy and bulk-phase enzymology to propose a mechanism for the resolution and extension of parallel G4 by telomerase. Binding is initiated by the RNA template of telomerase interacting with the G-quadruplex; nucleotide addition then proceeds to the end of the RNA template. It is only through the large conformational change of translocation following synthesis that the G-quadruplex structure is completely unfolded to a linear product. Surprisingly, parallel G4 stabilization with either small molecule ligands or by chemical modification does not always inhibit G4 unfolding and extension by telomerase. These data reveal that telomerase is a parallel G-quadruplex resolvase. © 2020, eLife Sciences Publications Ltd. All rights reserved.

Subject(s)

Alexafluor 555
 Chloroform
 Diagnostic agent
 Dimethyl sulfoxide
 Dithiothreitol
 Edetic acid
 Guanine quadruplex
 Levodopa
 Magnesium chloride
 Polysorbate 20
 Potassium chloride
 Sodium tetraborate decahydrate buffer
 Telomerase
 Telomerase reverse transcriptase
 Triton x 100
 Unclassified drug
 Article
 Chromosomal parameters
 Circular dichroism
 Computer simulation
 Controlled study
 Crystal structure
 DNA binding
 Fluorescence microscopy
 Fluorescence resonance energy transfer
 Fractional anisotropy
 Gene frequency
 Genetic association
 Genomic instability
 High performance liquid chromatography
 Human
 Human cell
 Image analysis
 Liquid chromatography-mass spectrometry
 Molecular diagnosis
 Molecular mechanics
 Molecular model
 Protein unfolding
 Telomerase activity assay
 Telomere
 Telomere length
 Thermostability
 Ultraviolet visible spectroscopy

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URI

https://hdl.handle.net/20.500.14740/4517

Collections

Scopus: Year 1983-2021

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