Transcription initiation of the copy-number control and better-than-random segregation genes broad-host-range low-copy-number plasmid pSM19035 are subjected to repression by autoregulated pSM19035-encoded ω product in Bacillus subtilis cells. The promoters copS ( P cop1 cop2), δ δ), ω) have been mapped. These embedded a set either seven copies 7-bp direct repeat or block consisting two repeats one inverted repeat; blocks present three times. cooperative binding protein on cop1, cop2, δ,...

Cytological and genetic evidence suggests that the Bacillus subtilis DNA uptake machinery localizes at a single cell pole takes up single-stranded (ss) DNA. The integration of homologous donor into recipient chromosome requires RecA, while plasmid establishment, which is independent least RecO RecU. RecA RecN colocalize polar machinery, from forms filamentous structures, termed threads, in presence chromosomal We show transformation follows distinct pathways. In absence DNA, RecU accumulated...

ABSTRACT Adenovirus is one of the most complex icosahedral, nonenveloped viruses. Even after its structure was solved at near-atomic resolution by both cryo-electron microscopy and X-ray crystallography, location minor coat proteins still a subject debate. The elaborated capsid architecture product correspondingly assembly process, about which many aspects remain unknown. Genome encapsidation involves concerted action five virus proteins, proteolytic processing protease needed to prime...

Yin Yang 1 (YY1) is a transcription factor regulating proliferation and differentiation involved in cancer development. Oligomers of recombinant YY1 have been observed before, but their structure DNA binding properties are not well understood. Here we find that assembles several homo-oligomeric species built from the association bell-shaped dimer, process characterized by electron microscopy. Moreover, self-association also occurs vivo using bimolecular fluorescence complementation....

Bacillus subtilis RecU protein is involved in homologous recombination, DNA repair, and chromosome segregation. Purified binds preferentially to three- four-strand junctions when compared single-stranded (ssDNA) double-stranded (dsDNA) (≈10- ≈40-fold lower efficiency, respectively). cleaves mobile four-way but fails cleave a linear dsDNA with putative cognate site, finding consistent similar genetic defect observed for genes classified within the ε epistatic group (namely ruv A, rec D, U)....

Two extracellular proteases from Staphylococcus hyicus subsp. hyicus, ShpI and ShpII, have been characterized. is a neutral metalloprotease with broad substrate specificity; the gene has cloned sequenced. characterized here, mainly produced in late logarithmic growth phase contrast to ShpI, which stationary phase. ShpII was purified culture medium of S. by ammonium sulfate precipitation DEAE-Sepharose chromatography. The molecular mass, estimated sodium dodecyl sulfate-polyacrylamide gel...

The ζε module consists of a labile antitoxin protein, ε, which in dimer form (ε2) interferes with the action long-living monomeric ζ phosphotransferase toxin through protein complex formation. Toxin ζ, inhibits cell wall biosynthesis and may be bactericide nature, at or near physiological concentrations induces reversible cessation Bacillus subtilis proliferation (protective dormancy) by targeting essential metabolic functions followed propidium iodide (PI) staining fraction (20–30%)...

Holliday junction (HJ) resolution is a fundamental step for completion of homologous recombination. HJ resolving enzymes (resolvases) distort the structure upon binding and prior cleavage, raising possibility that reactivity enzyme can be affected by particular geometry topology at junction. Here, we employed DNA origami nano-scaffold in which each arm was tethered through base-pair hybridization, allowing us to make core either flexible or inflexible adjusting length arms. Both junctions...

Streptococcus pyogenes pSM19035-encoded epsilon (10.7 kDa) and zeta (32.4 proteins are necessary to secure stable plasmid inheritance in bacteria, with acting as toxin that kills plasmid-deprived cells an antitoxin neutralises the activity of zeta. The co-purify a complex that, according analytical ultracentrifugation gel filtration, exists epsilon2zeta2 heterotetramer solution. Co-crystals contain 1:1 molar ratio. Unfolding studies monitoring circular dichroic fluorescence changes show...

Genetic evidence suggests that the SPP1-encoded gene <i>35</i> product (G<i>35</i>P) is essential for phage DNA replication. Purified G<i>35</i>P binds single-strand (ssDNA) and double-strand (dsDNA) specifically interacts with replicative helicase G<i>40</i>P SSB protein G<i>36</i>P. promotes joint molecule formation between a circular ssDNA homologous linear dsDNA an tail. Joint requires metal ion but independent of nucleotide cofactor. molecules formed during these reactions contain...

ABSTRACT Bacillus subtilis mutants classified within the ε ( ruvA , Δ ruvB recU and recD ) η (Δ recG epistatic groups, in an otherwise rec + background, render cells impaired chromosomal segregation. A less-pronounced segregation defect recA sms radA was observed. The repair deficiency of addAB recO recR recH recS subA did not correlate with a defect. sensitivity group to DNA-damaging agents correlates ongoing DNA replication at time exposure agents. mutations partially suppress cells. but...

Subsaturating amounts of Bacillus subtilis SsbA, independently the order addition, partially inhibit single-stranded DNA-dependent dATPase activity RecA. This negative effect is fully overcome when a substoichiometric amount RecO added. SsbA added prior to RecA does not stimulate dATP-dependent DNA strand exchange activity; however, after it enhances extent exchange. The addition stimulates RecA-mediated joint molecule formation, although limits accumulation final recombination products....

Abstract Bacillus subtilis RecA is important for spore resistance to DNA damage, even though spores contain a single non-replicating genome. We report that inactivation of or its accessory factors, RecF, RecO, RecR and RecX, drastically reduce survival mature dormant ultrahigh vacuum desiccation ionizing radiation induce strand (ss) nicks double-strand breaks (DSBs). The presence non-cleavable LexA renders less sensitive DSBs, impaired in DSB recognition end-processing show sensitivities...

The histone-like protein HU is a conserved nucleoid-associated that involved in the maintenance of bacterial chromosome architecture. It only known Streptococcus pneumoniae, but it has not been studied. pneumococcal gene encoding this protein, hlp, shown herein to be essential for cell viability. Its disruption was possible either when duplicated and its expression induced from PZn promoter, or hlp cloned into plasmid under control inducible Pmal promoter. In vitro assays indicated shows...

Complex viruses that encode their own initiation proteins and subvert the host's elongation apparatus have provided valuable insights into DNA replication. Using purified bacteriophage SPP1 Bacillus subtilis proteins, we reconstituted a rolling circle replication system recapitulates genetically defined protein requirements. Eleven are required: phage-encoded helicase (G40P), loader (G39P), origin binding (G38P) G36P single-stranded DNA-binding (SSB); host-encoded PolC DnaE polymerases,...