Single-molecule visualization of DNA G-quadruplex formation in live cells.

Nature Chemistry
Marco Di AntonioShankar Balasubramanian

Abstract

Substantial evidence now exists to support that formation of DNA G-quadruplexes (G4s) is coupled to altered gene expression. However, approaches that allow us to probe G4s in living cells without perturbing their folding dynamics are required to understand their biological roles in greater detail. Herein, we report a G4-specific fluorescent probe (SiR-PyPDS) that enables single-molecule and real-time detection of individual G4 structures in living cells. Live-cell single-molecule fluorescence imaging of G4s was carried out under conditions that use low concentrations of SiR-PyPDS (20 nM) to provide informative measurements representative of the population of G4s in living cells, without globally perturbing G4 formation and dynamics. Single-molecule fluorescence imaging and time-dependent chemical trapping of unfolded G4s in living cells reveal that G4s fluctuate between folded and unfolded states. We also demonstrate that G4 formation in live cells is cell-cycle-dependent and disrupted by chemical inhibition of transcription and replication. Our observations provide robust evidence in support of dynamic G4 formation in living cells.

References

Jul 5, 2001·Proceedings of the National Academy of Sciences of the United States of America·C SchaffitzelA Plückthun
Dec 3, 2003·Proceedings of the National Academy of Sciences of the United States of America·Liming YingShankar Balasubramanian
Jan 31, 2007·Journal of the American Chemical Society·Anne De CianDavid Monchaud
Jan 8, 2008·Nature Methods·Makio TokunagaKumiko Sakata-Sogawa
Nov 4, 2008·Journal of the American Chemical Society·Raphaël RodriguezShankar Balasubramanian
Feb 7, 2012·Nature Chemical Biology·Raphaël RodriguezStephen P Jackson
Jan 25, 2013·Nature Chemistry·Gražvydas LukinavičiusKai Johnsson
Feb 21, 2013·Nature Chemistry·Giulia BiffiShankar Balasubramanian
Jun 10, 2015·Journal of the American Chemical Society·Aurélien LaguerreDavid Monchaud
Jul 21, 2015·Nature Biotechnology·Vicki S ChambersShankar Balasubramanian
Sep 13, 2016·Nature Genetics·Robert Hänsel-HertschShankar Balasubramanian
Oct 8, 2016·Methods : a Companion to Methods in Enzymology·Jean-Yves TinevezKevin W Eliceiri
Feb 23, 2017·Nature Reviews. Molecular Cell Biology·Robert Hänsel-HertschShankar Balasubramanian
Feb 18, 2018·Angewandte Chemie·Xiu-Cai ChenJia-Heng Tan
May 10, 2018·Biophysical Journal·Aleks PonjavicSteven F Lee
May 9, 2019·Biomedical Optics Express·Aleks PonjavicDavid Klenerman

❮ Previous
Next ❯

Citations

Aug 26, 2020·Chemistry : a European Journal·Yanwei CaoRenjun Pei
Oct 13, 2020·Nucleic Acids Research·Ke-Wei ZhengZheng Tan
Oct 22, 2020·DNA Research : an International Journal for Rapid Publication of Reports on Genes and Genomes·Saniya M JavadekarSathees C Raghavan
Dec 9, 2020·Nucleic Acids Research·Kevin LiStephen Neidle
Jan 25, 2021·BMC Genomics·Václav BrázdaJean-Louis Mergny
Jan 24, 2021·Nature Chemical Biology·Haizhou Zhao, Chun Kit Kwok
Dec 29, 2020·Chembiochem : a European Journal of Chemical Biology·Pallabi SenguptaSubhrangsu Chatterjee
Nov 25, 2020·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Ahmed Abdullah Ahmed, Stephen Neidle
Dec 29, 2020·Microbiology and Molecular Biology Reviews : MMBR·Lucie Poggi, Guy-Franck Richard
Feb 13, 2021·International Journal of Biological Macromolecules·Xiaoxuan XiangXinhua Guo
Jan 16, 2021·Chembiochem : a European Journal of Chemical Biology·Mikayel AznauryanVictoria Birkedal
Mar 7, 2021·International Journal of Molecular Sciences·Diana SantosFrancesca M Pisani
Dec 12, 2020·Trends in Biochemical Sciences·Leïla DumasStefania Millevoi
Mar 28, 2021·Nucleic Acids Research·Kaixin LyuChun Kit Kwok
Apr 4, 2021·Genes·Mohammad MahtabFrancesca M Pisani
May 1, 2021·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Peter Jonas Wickhorst, Heiko Ihmels
Apr 29, 2021·Journal of Medicinal Chemistry·Meysam KhosravifarsaniBrigitte Guérin
Apr 15, 2021·Chemistry : a European Journal·Raj PaulJyotirmayee Dash
May 9, 2021·Genome Biology·Jiazhen ShenShankar Balasubramanian
May 12, 2021·Journal of Natural Products·Alexander S TikhomirovAndrey E Shchekotikhin
May 13, 2021·Nucleic Acids Research·Enrico CadoniAnnemieke Madder
May 20, 2021·Nucleic Acids Research·Phong Lan Thao TranVincent Croquette
Jun 30, 2021·Nature Chemistry·Xiaoyun ZhangShankar Balasubramanian
Jul 22, 2021·Nucleic Acids Research·Mo YangJohn S Schneekloth
Jul 14, 2021·Nucleic Acids Research·Jenna RobinsonMarco Di Antonio
Jul 25, 2021·International Journal of Molecular Sciences·Adriana VolnáJiří Červeň
Nov 19, 2020·The Journal of Physical Chemistry Letters·Davide AvaglianoLeticia González
Sep 22, 2020·Journal of the American Chemical Society·Yutaro TsubonoHiroshi Sugiyama
Oct 20, 2020·Journal of the American Chemical Society·Jim-Marcel KnopRoland Winter

❮ Previous
Next ❯

Methods Mentioned

BETA
immunoprecipitation
fluorescence imaging
circular dichroism
FRET
confocal microscopy
ChIP-Seq
fluorescence microscopy
fluorescence titrations

Software Mentioned

Prism
TrackMate
ImageJ
Cary Kinetics

Related Concepts

Related Feeds

Adhesion Molecules in Health and Disease

Cell adhesion molecules are a subset of cell adhesion proteins located on the cell surface involved in binding with other cells or with the extracellular matrix in the process called cell adhesion. In essence, cell adhesion molecules help cells stick to each other and to their surroundings. Cell adhesion is a crucial component in maintaining tissue structure and function. Discover the latest research on adhesion molecule and their role in health and disease here.

Cell Adhesion Molecules in the Brain

Cell adhesion molecules found on cell surface help cells bind with other cells or the extracellular matrix to maintain structure and function. Here is the latest research on their role in the brain.