Direct visualization of nucleolar G-quadruplexes in live cells by using a fluorescent light-up probe

Zhang, Suge; Sun, Hongxia; Chen, Hongbo; Li, Qian; Guan, Aijiao; Wang, Lixia; Shi, Yulei; Xu, Shujuan; Liu, Meirong; Tang, Yalin

doi:10.1016/j.bbagen.2018.01.022

Cited by 47 publications

(36 citation statements)

References 41 publications

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“…ThT-Pyridostatin (PDS) competition. PDS is a G-quadruplex stabilizer that binds with greater affinity than ThT to G-quadruplexes and can displace ThT from G-quadruplexes [20]. The results here show that G-quadruplex-induced ThT fluorescence of GQes3 is attenuated by addition of PDS, consistent with displacement of ThT from G-quadruplexes by PDS.…”

Section: Methodssupporting

confidence: 72%

Profusion of G-quadruplexes on both subunits of metazoan ribosomes

et al. 2019

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Mammalian and bird ribosomes are nearly twice the mass of prokaryotic ribosomes in part because of their extraordinarily long rRNA tentacles. Human rRNA tentacles are not fully observable in current three-dimensional structures and their conformations remain to be fully resolved. In previous work we identified sequences that favor G-quadruplexes in silico and in vitro in rRNA tentacles of the human large ribosomal subunit. We demonstrated by experiment that these sequences form G-quadruplexes in vitro. Here, using a more recent motif definition, we report additional G-quadruplex sequences on surfaces of both subunits of the human ribosome. The revised sequence definition reveals expansive arrays of potential G-quadruplex sequences on LSU tentacles. In addition, we demonstrate by a variety of experimental methods that fragments of the small subunit rRNA form G-quadruplexes in vitro. Prior to this report rRNA sequences that form G-quadruplexes were confined to the large ribosomal subunit. Our combined results indicate that the surface of the assembled human ribosome contains numerous sequences capable of forming G-quadruplexes on both ribosomal subunits. The data suggest conversion between duplexes and G-quadruplexes in response to association with proteins, ions, or other RNAs. In some systems it seems likely that the integrated population of RNA G-quadruplexes may be dominated by rRNA, which is the most abundant cellular RNA.

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Section: Methodssupporting

confidence: 72%

Profusion of G-quadruplexes on both subunits of metazoan ribosomes

et al. 2019

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“…Following the in vitro evidence of the high selectivity of CQ4 for parallel DNA G4 structures, we treated cells with RNase to confirm the nature of the main binding target of the compound (Figure B, upper panel). Thioflavin‐T (ThT) was used as control since it is also intracellularly fluorescent and known to bind RNA G4s making it highly sensitive to RNase treatment (Figure B, lower panel) . While RNase treatment did not modify CQ4 nucleolar staining, the nucleolar ThT signal was reduced by the presence of the enzyme, indicating the ability of CQ4 to preferentially target DNA G4 structures.…”

Section: Resultsmentioning

confidence: 99%

“…Thioflavin-T (ThT) was used as control since it is also intracellularly fluorescent and known to bind RNAG4s making it highly sensitive to RNase treatment ( Figure 9B,l ower panel). [17] While RNase treatment did not modify CQ4 nucleolar staining,t he nucleolar ThTs ignal was reduced by the presence of the enzyme, indicating the ability of CQ4 to preferentially target DNAG4 structures.D Nase treatment was not performed due to the apparent inability of this enzyme to reach the nucleoli. [16c] Instead, DNAG 4b inding was confirmed by using the wellknown G4-binder Phen-DC 3 (Figures 9C and Supporting Information, Figure S47).…”

Section: Cellular Imagingmentioning

confidence: 89%

A Light‐up Logic Platform for Selective Recognition of Parallel G‐Quadruplex Structures via Disaggregation‐Induced Emission

et al. 2019

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The design of turn‐on dyes with optical signals sensitive to the formation of supramolecular structures provides fascinating and underexplored opportunities for G‐quadruplex (G4) DNA detection and characterization. Here, we show a new switching mechanism that relies on the recognition‐driven disaggregation (on‐signal) of an ultrabright coumarin‐quinazoline conjugate. The synthesized probe selectively lights‐up parallel G4 DNA structures via the disassembly of its supramolecular state, demonstrating outputs that are easily integrable into a label‐free molecular logic system. Finally, our molecule preferentially stains the G4‐rich nucleoli of cancer cells.

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“…It has been reported that ThT exhibits high selectivity for G-quadruplex structures in vitro (28–31). However, in living cells, ThT mainly stains the nucleoli (32), making it impossible to detect the G-quadruplex DNA in chromatin. Nevertheless, the exciting properties of ThT inspire us to develop new benzothiazole family compounds (detailed syntheses and characterization are given in the Supplementary Data) in the hope of obtaining an ideal probe for the direct detection of G-quadruplex DNA in living cells.…”

Section: Introductionmentioning

confidence: 99%

Real-time monitoring of DNA G-quadruplexes in living cells with a small-molecule fluorescent probe

Zhang

Sun

Wang

et al. 2018

Nucleic Acids Research

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123

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G-quadruplex DNA has been viewed as a prospective anti-cancer target owing to its potential biological relevance. Real-time monitoring of DNA G-quadruplex structures in living cells can provide valuable insights into the relationship between G-quadruplex formation and its cellular consequences. However, the probes capable of detecting DNA G-quadruplexes in living cells are still very limited. Herein, we reported a new fluorescent probe, IMT, for real-time visualization of DNA G-quadruplex structures in living cells. Using IMT as a fluorescent indicator, the quantity changes of DNA G-quadruplex at different points in time during continuous cellular progression responding to Aphidicolin and Hydroxyurea treatment have been directly visualized. Our data demonstrate that IMT will be a valuable tool for exploring DNA G-quadruplexes in live cells. Further application of IMT in fluorescence imaging may reveal more information on the roles of DNA G-quadruplexes in biological systems.

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Direct visualization of nucleolar G-quadruplexes in live cells by using a fluorescent light-up probe

Cited by 47 publications

References 41 publications

Profusion of G-quadruplexes on both subunits of metazoan ribosomes

Profusion of G-quadruplexes on both subunits of metazoan ribosomes

A Light‐up Logic Platform for Selective Recognition of Parallel G‐Quadruplex Structures via Disaggregation‐Induced Emission

Real-time monitoring of DNA G-quadruplexes in living cells with a small-molecule fluorescent probe

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