RecQ family helicases are critical for maintaining genomic integrity. Many RecQ family helicases not only unwind duplex, and other more complicated DNA structures, but also possess, interestingly, DNA annealing (strand pairing) activity. Here, we systematically investigated the DNA annealing properties of RECQ5β by measuring DNA annealing kinetics, equilibrium DNA binding, and kinetics of dissociation from ssDNA. RECQ5β catalyzed DNA annealing most efficiently when the enzyme molecules covered approximately 40%-50% of the DNA strand, in the absence or presence of different nucleotide cofactors (AMPPNP, ATPγS, or ADP) under our buffer conditions. A comparative study with RECQ5β1-662 confirmed that the C-terminal region of RECQ5β was essential for its high DNA annealing activity. These results contribute to our understanding of the mechanism of DNA annealing catalyzed by RecQ family helicases.
Dual-layer Metallic grating(DMG)structures as surface-enhanced Raman scattering(SERS)substrates are studied using benzenethiol as the probe analyte.The DMG structure consists of a SiO2 grating and 100-nm-thick gold coating layers.An enhancement factor of 105 is achieved by optimizing the SiO 2 grating height within the range from 165 to 550 nm.The enhancement factor dependence on the SiO2 grating height is due to the surface plasmon excitation,which is dependent on the polarization of the incident light,and confirmed by finite difference time domain simulations.This study demonstrates the advantages of high uniformity,reproducibility and sensitivity in the DMG structures for SERS applications.
With atomic force microscopy (AFM) we systematically studied the DNA condensations on mica surfaces induced by multivalent cation spermidine. The pattern of the DNA condensates is a flat single layer, with a core in the centre and DNA wrapping around it at high density. We assume this to be a two-dimensional condensation of free coiled DNA onto negatively charged mica surfaces by the multivalent cation. The DNA molecules condense on mica surfaces via a pathway different from the formation of toroids, rods or globules in bulk solutions. We give an explanation to why toroid structures are difficult to be observed by AFM, and further discuss the relationship between DNA condensations in solutions and on mica surfaces. The present work will be helpful for understanding the behaviors of DNA on charged surfaces, which might be significantly different from that in solutions.
ZHANG XingHua HOU XiMiao JI Chao LIMing DOU ShuoXing WANG PengYe
The DNA gyrase of Escherichia coli plays an essential role in the life of this microorganism.It is unique among all topoisomerases because of its ability to introduce negative supercoils into DNA.This study investigated the single molecular interaction of E.coli gyrase with DNA using magnetic tweezers.The results showed that,in the absence of ATP,gyrase weakly binds the G and T segments.The stretched force of 0.7 pN can gradually destroy the binding,whereas that of 5.9 pN directly destroys it.Addition of high concentrations of norfloxacin enhances gyrase binding to both segments,making them adapt to 5.9 pN.DNA gyrase reduces the plectonemic dimension,which was determined by the bacterial enzyme and not by the pull force.Moreover,it has different affinities for positive supercoils,which it prefers,and negative supercoils.The time distribution of the dissociation of gyrase from DNA has a double-exponential form.We herein propose a model to explain this distribution and compare the results with those of other models.
The effect of the median selective solution on the lamellar,spherical and gyroid structures is studied. The self-consistent field equations of the diblock copolymer solution are solved by using the reciprocal space method. It is shown that the spherical and gyroid phases have the lowest free energy in the certain range of the solution concentration. Furthermore,the phase diagram of the ordered structures in the diblock copolymer solution with the median selective solvent is calculated,which is consistent with the experimental results.
