UHRF1 plays a role in maintaining DNA methylation in mammalian cells

Epigenetic inheritance in mammals relies in part on robust propagation of DNA methylation patterns throughout development. We show that the protein UHRF1 (ubiquitin-like, containing PHD and RING finger domains 1), also known as NP95 in mouse and ICBP90 in human, is required for maintaining DNA methylation. UHRF1 colocalizes with the maintenance DNA methyltransferase protein DNMT1 throughout S phase. UHRF1 appears to tether DNMT1 to chromatin through its direct interaction with DNMT1. Furthermore UHRF1 contains a methyl DNA binding domain, the SRA (SET and RING associated) domain, that shows strong preferential binding to hemimethylated CG sites, the physiological substrate for DNMT1. These data suggest that UHRF1 may help recruit DNMT1 to hemimethylated DNA to facilitate faithful maintenance of DNA methylation.     

Regulation of replication fork progression through histone supply and demand

DNA replication in eukaryotes requires nucleosome disruption ahead of the replication fork and reassembly behind. An unresolved issue concerns how histone dynamics are coordinated with fork progression to maintain chromosomal stability. Here, we characterize a complex in which the human histone chaperone Asf1 and MCM2-7, the putative replicative helicase, are connected through a histone H3-H4 bridge. Depletion of Asf1 by RNA interference impedes DNA unwinding at replication sites, and similar defects arise from overproduction of new histone H3-H4 that compromises Asf1 function. These data link Asf1 chaperone function, histone supply, and replicative unwinding of DNA in chromatin. We propose that Asf1, as a histone acceptor and donor, handles parental and new histones at the replication fork via an Asf1-(H3-H4)-MCM2-7 intermediate and thus provides a means to fine-tune replication fork progression and histone supply and demand.     

Raman-assisted crystallography reveals end-on peroxide intermediates in a nonheme iron enzyme

Iron-peroxide intermediates are central in the reaction cycle of many iron-containing biomolecules. We trapped iron(III)-(hydro)peroxo species in crystals of superoxide reductase (SOR), a nonheme mononuclear iron enzyme that scavenges superoxide radicals. X-ray diffraction data at 1.95 angstrom resolution and Raman spectra recorded in crystallo revealed iron-(hydro)peroxo intermediates with the (hydro)peroxo group bound end-on. The dynamic SOR active site promotes the formation of transient hydrogen bond networks, which presumably assist the cleavage of the iron-oxygen bond in order to release the reaction product, hydrogen peroxide.     

A cultured greigite-producing magnetotactic bacterium in a novel group of sulfate-reducing bacteria

Magnetotactic bacteria contain magnetosomes--intracellular, membrane-bounded, magnetic nanocrystals of magnetite (Fe(3)O(4)) or greigite (Fe(3)S(4))--that cause the bacteria to swim along geomagnetic field lines. We isolated a greigite-producing magnetotactic bacterium from a brackish spring in Death Valley National Park, California, USA, strain BW-1, that is able to biomineralize greigite and magnetite depending on culture conditions. A phylogenetic comparison of BW-1 and similar uncultured greigite- and/or magnetite-producing magnetotactic bacteria from freshwater to hypersaline habitats shows that these organisms represent a previously unknown group of sulfate-reducing bacteria in the Deltaproteobacteria. Genomic analysis of BW-1 reveals the presence of two different magnetosome gene clusters, suggesting that one may be responsible for greigite biomineralization and the other for magnetite.     

Tracking solar gravity modes: the dynamics of the solar core

Solar gravity modes have been actively sought because they directly probe the solar core (below 0.2 solar radius), but they have not been conclusively detected in the Sun because of their small surface amplitudes. Using data from the Global Oscillation at Low Frequency instrument, we detected a periodic structure in agreement with the period separation predicted by the theory for gravity dipole modes. When studied in relation to simulations including the best physics of the Sun determined through the acoustic modes, such a structure favors a faster rotation rate in the core than in the rest of the radiative zone.     

Chromosomal gene transfer in Spiroplasma citri

The study of resistance marker rearrangement in Spiroplasma citri mutants provides evidence of transfer of chromosomal information followed by recombination. This is the first report of such a transfer in vivo in the mollicutes--that is, in the smallest self-replicating organisms. The double-resistant phenotypes obtained are stable even without selection pressure. The mechanism of gene transfer is insensitive to deoxyribonuclease, requires contact, and possibly, areas of fusion of the cell membranes; it shares properties with the transfer by protoplast fusion in Gram-positive bacteria. The extensive degenerative evolution of mollicutes has retained, in S. citri, bacterial functions of chromosomal transfer and recombination.     

Recurrent selection for resistance to Verticillium dahliae in pepper

Summary Two cycles of recurrent selection were performed in a population including 7 small fruited varieties of pepper (Capsicum annuum) with low resistance level to Verticillium and resistance to other soilborne pathogens. The collaboration of 7 plant breeders in this work and the use of 2 artificial methods of inoculation allowed screening of numerous plants for resistance to V. dahliae under different conditions. The population was simultaneously improved for resistance to V. dahliae and for resistance to Phytophthora capsici, and at each cycle, intercrosses were performed between all these plants. The general level of resistance to V. dahliae increased continuously during the 2 cycles of seles of selection. Selfed progenies of plants from the 2 cycles displayed a higher resistance than the original parents, whatever the method of inoculation used. This gave evidence of transgressions for resistance. Crosses involving parents that did not seem resistant by themselves also produced transgressive lines, indicating that resistance genes were present but not active in these varieties. The transgressive lines with polygenic resistance to V. dahliae were included into a further breeding program to introduce genes of agronomic value.     

Breeding strategy in foxtail millet,Setaria italica (L.P. Beauv.), following interspecific hybridization

Summary An interspecific cross between foxtail millet Setaria italica and its wild relative Setaria viridis was undertaken in order to introduce from the latter triazine resistance in the cultivated plant. Four backcross generations obtained with S. italica as recurrent parent were studied. Results show that only two backcross generations associated with selection are enough to eliminate weedy characters and to return to the cultivated type. S. viridis could thus be an interesting source for improvement of foxtail millet without a complex and time consuming breeding strategy.     

Variation and covariation of silage maize digestibility estimated from digestion trials with sheep

Summary Variation and covariation for agronomic and digestibility traits of silage maize are reported from a compilation of 22 years of experiments with standard sheep. Genotype effects of DOM and DCF were highly significant, even when genotypes were nested in earliness groups or brown-midrib hybrids discarded (Table 2). The genetic variance of crude fiber content was low, but the variance of the DCF was high. The genetic variance of DOM was about 4 times lower than genetic variance of DCF, but broad sense heritability of DOM was higher because of lower residual variance (Table 3). Genetic correlations between grain or crude fiber content and DOM had similar absolute values, 0.65, so each of these two traits was an important but not the unique determinant of silage maize quality. There was no correlation between DCF and grain or crude fiber content. Yield was not related to DOM or DCF within each group of earliness, allowing some quality improvement without agronomic drift (Table 4). Except for late hybrids, most of DOM differences between groups of earliness came from lowering of minimum value, while maximum values were similar. It was the contrary for DCF, with similar minimum values for all groups (Table 5). There was no obvious correlation between year of registration of hybrids and DOM or DCF, but extra new variation seemed obtained only for low values (Figs 1, 2; Table 6). IVDOM according to the APC process was a poor predictor of DOM, especially when brown-midrib hybrids and earliness effects were discarded; but because heritability of this trait was similar to DOM heritability, such enzymatic processes could probably be used to avoid drift towards poor DOM with hybrids bred for higher stalk strength.Abbreviations APC Amylase Pepsine Cellulase solubility - DDM sheep digestibility of dry matter - DOM sheep digestibility of organic matter - DCF sheep digestibility of crude fiber - DM dry matter - IVDCW in vitro digestibility of cell wall - IVDDM in vitro digestibility of dry matter - IVDOM in vitro digestibility of organic matter - NIRS near infra-red reflectance spectroscopy - NDF neutral detergent fiber - bm₁-brown-midrib-1 allele, bm₃-brown midrib-3 allele