Geometry-Based Self-Assembly of Histone-DNA Nanostructures at Single-Nucleotide Resolution.

Histones are basic protein monomers capable of interacting with DNA, providing the mechanism of DNA compaction inside the cell nucleus. The well-ordered assembly process of histone and DNA is a potential candidate as the approach for building DNA-protein nanostructures. Here, utilizing the sequence-independent histone-DNA interaction, we present an approach to self-assemble histones and single-stranded DNA (ssDNA) to form well-defined histone-DNA (sHD) nanoparticles and their multidimensional cross-linked complexes (cHD). By using various molecular biology and microscopy techniques, we elucidate the structure of these complexes, and we show that they are formed at carefully controlled conditions of temperature, ionic strength, concentration, and incubation time. We also demonstrate using a set of ssDNA molecular rulers and a geometric accommodation model that the assembly of sHD and cHD particles proceeds with precise geometry so that the number of ssDNA in these particles can be programmed by the length of ssDNA. We further show that the formation of cHD amplifies the effect of the length of ssDNA on the self-assembly, allowing for distinguishing ssDNA of different lengths at single nucleotide resolution. We envision that our geometry-directed approach of self-assembling histone-DNA nanostructures and the fundamental insights can serve as a structural platform to advance building precisely ordered DNA-protein nanostructures.

Entangled polymer dynamics beyond reptation.

Macroscopic properties of polymers arise from microscopic entanglement of polymer chains. Entangled polymer dynamics have been described theoretically by time- and space-averaged relaxation modes of single chains occurring at different time and length scales. However, theoretical and experimental studies along this framework provide oversimplified picture of spatiotemporally heterogeneous polymer dynamics. Characterization of entangled polymer dynamics beyond this paradigm requires a method that enables to capture motion and relaxation occurring in real space at different length and time scales. Here we develop new single-molecule characterization platform by combining super-resolution fluorescence imaging and recently developed single-molecule tracking method, cumulative-area tracking, which enables to quantify the chain motion in the length and time scale of nanometres to micrometres and milliseconds to minutes. Using linear and cyclic dsDNA molecules as model systems, our new method reveals chain-position-dependent motion of the entangled linear chains, which is beyond the scope of current theoretical framework.

Single-molecule diffusion and conformational dynamics by spatial integration of temporal fluctuations.

Single-molecule localization and tracking has been used to translate spatiotemporal information of individual molecules to map their diffusion behaviours. However, accurate analysis of diffusion behaviours and including other parameters, such as the conformation and size of molecules, remain as limitations to the method. Here, we report a method that addresses the limitations of existing single-molecular localization methods. The method is based on temporal tracking of the cumulative area occupied by molecules. These temporal fluctuations are tied to molecular size, rates of diffusion and conformational changes. By analysing fluorescent nanospheres and double-stranded DNA molecules of different lengths and topological forms, we demonstrate that our cumulative-area method surpasses the conventional single-molecule localization method in terms of the accuracy of determined diffusion coefficients. Furthermore, the cumulative-area method provides conformational relaxation times of structurally flexible chains along with diffusion coefficients, which together are relevant to work in a wide spectrum of scientific fields.

Complete genome sequence of Mycobacterium vaccae type strain ATCC 25954.

Mycobacterium vaccae is a rapidly growing, nontuberculous Mycobacterium species that is generally not considered a human pathogen and is of major pharmaceutical interest as an immunotherapeutic agent. We report here the annotated genome sequence of the M. vaccae type strain, ATCC 25954.