In this research, an unusually dimeric G-quadruplex of d（GGGTGGGTGGGTGGGT） （SI）, the potent nanomolar HIV-1 integrase inhibitor, was detected by nuclear magnetic resonance （NMR）. This result has been confirmed b...In this research, an unusually dimeric G-quadruplex of d（GGGTGGGTGGGTGGGT） （SI）, the potent nanomolar HIV-1 integrase inhibitor, was detected by nuclear magnetic resonance （NMR）. This result has been confirmed by electrospray ionization mass spectrometry （ESI-MS） and circular dichroism （CD）.展开更多
A simple, rapid, highly sensitive electrochem-ical sensor for potassium ion （K^＋） based on the confor-mationai change of DNA sequence containing guanine-rich segments is presented. In the presence of K^＋, guanine-...A simple, rapid, highly sensitive electrochem-ical sensor for potassium ion （K^＋） based on the confor-mationai change of DNA sequence containing guanine-rich segments is presented. In the presence of K^＋, guanine-rich DNA sequence folds to G-quadruplex structure, allowing a ferrocene tag to transfer electrons to the electrode. Gold nanoparticles （AuNPs）, which are self-assembled on the surface of a bare gold electrode by using 4-aminothio-phenol as a medium, offer a big surface area to immobilize a large number of aptamers and improve the sensitivity of the sensor. The square-wave voltammetry peak current increases with K^＋ concentration. The plots of peak current against K^＋ concentration and the logarithm of K^＋ con- centration are linear over the range from 0.1 to 1.0 mmol·L^-1 and from 1 to 30 mmol·L^-1, respectively. A lower detection limit of 0.1 mmol·L^-1 K^＋ is obtained for AuNPs-modified sensor, which greatly surpasses that （100 mmol·L^-1） of the sensor without AuNPs modification by three orders of magnitude. Thus, the sensor with AuNPs amplification is expected to open new opportunities for highly sensitive detection of other biomolecules in the future.展开更多
The Bloom helicase （BLM） gene product encodes a DNA helicase that functions in homologous recombination repair to prevent genomic instability. BLM is highly active in binding and unfolding G-quadruplexes （G4）, whi...The Bloom helicase （BLM） gene product encodes a DNA helicase that functions in homologous recombination repair to prevent genomic instability. BLM is highly active in binding and unfolding G-quadruplexes （G4）, which are non- canonical DNA structures formed by Hoogsteen base-pairing in guanine-rich sequences. Here we use single-molecule fluorescence resonance energy transfer （smFRET） to study the molecular mechanism of BLM-catalysed G4 unfolding and show that BLM unfolds G4 in two pathways. Our data enable us to propose a model in which the HRDC domain functions as a regulator of BLM, depending on the position of the HRDC domain of BLM in action： when HRDC binds to the G4 sequence, BLM may hold G4 in the unfolded state; otherwise, it may remain on the unfolded G4 transiently so that G4 can refold immediately.展开更多
Human telomeric G-quadruplex plays a crucial role in regulating the genome stability. Despite extensive studies on structures and kinetics of monomeric G-quadruplex, the interaction between G-quadruplexes is still in ...Human telomeric G-quadruplex plays a crucial role in regulating the genome stability. Despite extensive studies on structures and kinetics of monomeric G-quadruplex, the interaction between G-quadruplexes is still in debate. In this work,we employ magnetic tweezers to investigate the folding and unfolding kinetics of two contiguous G-quadruplexes in 100-mM K~＋buffer. The interaction between G-quadruplexes and the consequent effect on the kinetics of G-quadruplex are revealed. The linker sequence between G-quadruplexes is further found to play an important role in the interaction between two G-quadruplexes. Our results provide a high-resolution insight into kinetics of multimeric G-quadruplexes and genome stability.展开更多
基金the National Natural Science Foundation of China(No.20472009)the Research Fund for the Doctoral Program of Higher Education.
文摘In this research, an unusually dimeric G-quadruplex of d（GGGTGGGTGGGTGGGT） （SI）, the potent nanomolar HIV-1 integrase inhibitor, was detected by nuclear magnetic resonance （NMR）. This result has been confirmed by electrospray ionization mass spectrometry （ESI-MS） and circular dichroism （CD）.
基金financially supported by the National Natural Science Foundation of China(No.20903008)
文摘A simple, rapid, highly sensitive electrochem-ical sensor for potassium ion （K^＋） based on the confor-mationai change of DNA sequence containing guanine-rich segments is presented. In the presence of K^＋, guanine-rich DNA sequence folds to G-quadruplex structure, allowing a ferrocene tag to transfer electrons to the electrode. Gold nanoparticles （AuNPs）, which are self-assembled on the surface of a bare gold electrode by using 4-aminothio-phenol as a medium, offer a big surface area to immobilize a large number of aptamers and improve the sensitivity of the sensor. The square-wave voltammetry peak current increases with K^＋ concentration. The plots of peak current against K^＋ concentration and the logarithm of K^＋ con- centration are linear over the range from 0.1 to 1.0 mmol·L^-1 and from 1 to 30 mmol·L^-1, respectively. A lower detection limit of 0.1 mmol·L^-1 K^＋ is obtained for AuNPs-modified sensor, which greatly surpasses that （100 mmol·L^-1） of the sensor without AuNPs modification by three orders of magnitude. Thus, the sensor with AuNPs amplification is expected to open new opportunities for highly sensitive detection of other biomolecules in the future.
基金supported by the National Natural Science Foundation of China(Grant Nos.11674382,11574381,and 11574382)the Key Research Program of Frontier Sciences,Chinese Academy of Sciences(Grant No.QYZDJ-SSW-SYS014)
文摘The Bloom helicase （BLM） gene product encodes a DNA helicase that functions in homologous recombination repair to prevent genomic instability. BLM is highly active in binding and unfolding G-quadruplexes （G4）, which are non- canonical DNA structures formed by Hoogsteen base-pairing in guanine-rich sequences. Here we use single-molecule fluorescence resonance energy transfer （smFRET） to study the molecular mechanism of BLM-catalysed G4 unfolding and show that BLM unfolds G4 in two pathways. Our data enable us to propose a model in which the HRDC domain functions as a regulator of BLM, depending on the position of the HRDC domain of BLM in action： when HRDC binds to the G4 sequence, BLM may hold G4 in the unfolded state; otherwise, it may remain on the unfolded G4 transiently so that G4 can refold immediately.
基金supported by the National Natural Science Foundation of China(Grant Nos.11474346 and 11774407)the Key Research Program of Frontier Sciences,Chinese Academy of Sciences(Grant No.QYZDB-SSW-SLH045)the National Key Research and Development Program,China(Grant No.2016YFA0301500)
文摘Human telomeric G-quadruplex plays a crucial role in regulating the genome stability. Despite extensive studies on structures and kinetics of monomeric G-quadruplex, the interaction between G-quadruplexes is still in debate. In this work,we employ magnetic tweezers to investigate the folding and unfolding kinetics of two contiguous G-quadruplexes in 100-mM K~＋buffer. The interaction between G-quadruplexes and the consequent effect on the kinetics of G-quadruplex are revealed. The linker sequence between G-quadruplexes is further found to play an important role in the interaction between two G-quadruplexes. Our results provide a high-resolution insight into kinetics of multimeric G-quadruplexes and genome stability.
