It is well known that chromatin structure is highly sensitive to the ionic environment. However, the combined effects of a physiologically relevant mixed ionic environment of K+, Mg2+ and Na+, which are the main cations of the cell cytoplasm, has not been systematically investigated. We studied folding and self-association (aggregation) of recombinant 12-mer nucleosome arrays with 177 bp DNA repeat length in solutions of mixtures of K+ and Mg2+ or Na+ and Mg2+. In the presence of Mg2+, the addition of sodium ions promotes folding of array into 30-nm fibres, whereas in mixtures of K+ and Mg2+, potassium ions abrogate folding. We found that self-association of nucleosome arrays in mixed salt solutions is synergistically promoted by Mg2+ and monovalent ions, with sodium being slightly more efficient than potassium in amplifying the self-association. The results highlight the importance of a mixed ionic environment for the compaction of chromatin under physiological conditions and demonstrate the complicated nature of the various factors that determine and regulate chromatin compaction in vivo.

The accuracy and feasibility of computing the zenith tropospheric delays (ZTDs) from data of the European Center for Medium-Range Weather Forecasts (ECMWF) and the United States National Centers for Environmental Prediction (NCEP) are studied. The ZTDs are calculated from ECMWF/NCEP pressure-level data by integration and from the surface data with the Saastamoinen model method and then compared with the solutions measured from 28 global positioning system (GPS) stations of the Crustal Movement Observation Network of China (CMONOC) for 1 year. The results are as follows: (1) the error of the integration method is 1-3 cm less than that of the Saastamoinen model method. The agreement between the ECMWF ZTD and GPS ZTD is better than that between NCEP ZTD and GPS ZTD; (2) the bias and root mean square difference (RMSD), especially the latter, have a seasonal variation, and the RMSD decreases with increasing altitude while the variation with latitude is not obvious; and (3) when using the full horizontal resolution of 0.5°9 0.5°of the ECMWF meteorological data in place of a reduced 2.5°9 2.5°grid, the mean RMSD between GPS and ECMWF ZTD decreases by 4.5 mm. These results illuminated the accuracy and feasibility of computing the tropospheric delays and establishing the ZTD prediction model over China for navigation and positioning with ECMWF and NCEP data.Keywords GPS Á Zenith tropospheric delay (ZTD) Á CMONOC Á ECMWF Á NCEP Abbreviations CMONOCThe crustal movement observation network of China CDAS Climate data assimilation system ECMWF

Histone lysine methylations have primarily been linked to selective recruitment of reader or effector proteins that subsequently modify chromatin regions and mediate genome functions. Here, we describe a divergent role for histone H4 lysine 20 mono-methylation (H4K20me1) and demonstrate that it directly facilitates chromatin openness and accessibility by disrupting chromatin folding. Thus, accumulation of H4K20me1 demarcates highly accessible chromatin at genes, and this is maintained throughout the cell cycle. In vitro, H4K20me1-containing nucleosomal arrays with nucleosome repeat lengths (NRL) of 187 and 197 are less compact than unmethylated (H4K20me0) or trimethylated (H4K20me3) arrays. Concordantly, and in contrast to trimethylated and unmethylated tails, solid-state NMR data shows that H4K20 mono-methylation changes the H4 conformational state and leads to more dynamic histone H4-tails. Notably, the increased chromatin accessibility mediated by H4K20me1 facilitates gene expression, particularly of housekeeping genes. Altogether, we show how the methylation state of a single histone H4 residue operates as a focal point in chromatin structure control. While H4K20me1 directly promotes chromatin openness at highly transcribed genes, it also serves as a stepping-stone for H4K20me3-dependent chromatin compaction.

Telomeres, the ends of eukaryotic chromosomes, play pivotal roles in ageing and cancer and are targets of DNA damage and response. However, little is known about the structure and organization of telomeric chromatin at the molecular level. We used electron microscopy and single-molecule magnetic tweezers to characterize well-defined telomeric chromatin fibers of kilobasepair length. The cryo-EM structure of the compact telomeric tetranucleosome revealed a novel columnar folding, unusually short nucleosome repeat length of ~132bp and the role of the histone N-terminal tails in stabilizing this structure. This is the first near-high resolution structure of chromatin with a native DNA sequence. The columnar structure exposes the DNA, making them susceptible to DNA damage. The telomeric tetranucleosome also exists in an alternative well-defined state, with one nucleosome open, accessible to protein factors. This suggests that protein factors, which plays a role in maintaining telomeres, can bind to telomeric chromatin in its compact heterochromatic form. The features of the telomeric chromatin structure reveals important insights of significant relevance for telomere function in vivo that provides information on mechanisms of nucleosome recognition by chromatin factors

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The megabase-sized length of chromatin is highly relevant to the state of chromatin in vivo, where it is subject to a highly crowded environment and is organized in topologically associating domains of similar dimension. We developed an in vitro experimental chromatin model system reconstituted from T4 DNA (approximately 166 kbp) and histone octamers and studied the monomolecular compaction of this megabase-sized chromatin fiber under the influence of macromolecular crowding. We used single-molecule fluorescence microscopy and observed compaction in aqueous solutions containing poly(ethylene glycol) in the presence of monovalent (Na and K) and divalent (Mg) cations. Both DNA and chromatin demonstrated compaction under comparable conditions in the presence of poly(ethylene glycol) and Na or Mg salt. However, the mechanism of the compaction changed from a first-order phase transition for DNA to a continuous folding for megabase-sized chromatin fibers. A more efficient and pronounced chromatin compaction was observed in the presence of Na compared to K. A flow-stretching technique to unfold DNA and chromatin coils was used to gain further insight into the morphology of partially folded chromatin fibers. The results revealed a distribution of partially folded chromatin fibers. This variability is likely the result of the heterogeneous distribution of nucleosomes on the DNA chain. The packaging of DNA in the form of chromatin in the crowded nuclear environment appears essential to ensure gradual conformational changes of DNA.

Anionic macromolecular crowding promotes a very efficient compaction of chromatin fibers and self-assembly into micrometer-sized colloidal aggregates.

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