Starvation of cells for the DNA building block dTTP is strikingly lethal (thymineless death, TLD), and this effect observed in all organisms. The phenomenon, discovered some 60 years ago, widely used to kill anticancer therapies, but many questions regarding precise underlying mechanisms have remained. Here, we show first time that starvation precursor dGTP can E. coli a manner sharing features with TLD. accomplished by combining up-regulation cellular dGTPase deficiency guanine salvage...

Neisseria meningitidis is a leading cause of bacterial meningitis and sepsis worldwide an occasional meningococcal urethritis. When isolates are unavailable for surveillance or outbreak investigations, molecular characterization pathogens needs to be performed directly from clinical specimens, such as cerebrospinal fluid (CSF), blood, urine. However, genome sequencing specimens challenging because low high human DNA abundances. We developed selective whole-genome amplification (SWGA),...

The Escherichia coli dgt gene encodes a dGTP triphosphohydrolase whose detailed role still remains to be determined. Deletion of creates mutator phenotype, indicating that the dGTPase has fidelity role, possibly by affecting cellular dNTP pool. In present study, we have investigated structure Dgt protein at 3.1-Å resolution. One obtained structures revealed hexamer contained two molecules single-stranded DNA. presence DNA caused significant conformational changes in enzyme, including...

The larvicidal activity of Bacillus thuringiensis subsp. israelensis against dipteran larvae is determined by four major polypeptides the parasporal crystalline body produced during sporulation. Cyt1Aa shows lowest toxicity when used alone but most synergistic with any other proteins. sequence plasmid pBtoxis, which contains all toxin genes in this subspecies, revealed a new cyt-like coding named cyt1Ca. In addition to Cyt-like region, predicted Cyt1Ca contained an extra domain at C...

Summary The ATP‐bound form of the Escherichia coli DnaA replication initiator protein remodels chromosomal origin replication, oriC , to load replicative helicase. primary mechanism for regulating activity involves Hda and β clamp proteins, which act together dramatically stimulate intrinsic DNA‐dependent ATPase via a process termed Regulatory Inactivation DnaA. In addition hyperinitiation, strains lacking hda function also exhibit cold sensitive growth at 30°C. Strains impaired other...

Abstract Rollout of meningococcal serogroup A conjugate vaccine in Africa started 2010, aiming to eliminate meningitis outbreaks, belt countries. Since then, studies have been conducted, primarily using isolates, assess the impact on distribution strains region. Here, we implemented an innovative, culture-free whole-genome sequencing approach almost 400 clinical specimens collected between 2017 and 2019 from cases 6 African About 50% provided high-quality sequence data for comprehensive...

Compaction of the Escherichia coli nucleoid in cell's centre was associated with loss colony-forming ability; these effects were caused by induction Cyt1Aa, cytotoxic 27 kDa protein from Bacillus thuringiensis subsp. israelensis . Cyt1Aa-affected compaction nucleoids delayed but eventually more intense than chloramphenicol. The possibility that small, compact Cyt1Aa-expressing cells resulted DNA replication run-out and segregation following cell division ruled out measuring relative length....

Summary The Escherichia coli dGTP triphosphohydrolase (dGTPase) encoded by the dgt gene catalyses hydrolysis of to deoxyguanosine and triphosphate. recent discovery a mutator effect associated with deletion indicated participation in preventing mutagenesis. Here, we have investigated possible involvement facilitating thymine utilization through its ability provide intracellular deoxyguanosine, which is readily converted DeoD phosphorylase deoxyribose‐1‐phosphate, critical intermediate that...

The gene cyt1Aa is one of the genes in complex determining mosquito larvicidity Bacillus thuringiensis subsp. israelensis. Previous cloning Escherichia coli resulted a 48-bp addition upstream, encoding chimera. Here, was recloned without artifact, and its toxicity against Aedes aegypti larvae host E. cells retested.

ABSTRACT Our laboratory recently discovered that Escherichia coli cells starved for the DNA precursor dGTP are killed efficiently (dGTP starvation) in a manner similar to described thymineless death (TLD). Conditions specific starvation can be achieved by depriving an E. optA1 gpt strain of purine nucleotide hypoxanthine (Hx). To gain insight into mechanisms underlying starvation, we conducted genome-wide gene expression analyses actively growing subjected deprivation increasing periods. The...

Insecticidal crystal proteins of Bacillus thuringiensis belong to two unrelated toxin families: receptor-specific Cry toxins against insects and Cyt that lyse a broad range cells, including bacteria, via direct binding phospholipids. A new cyt-like open reading frame (cyt1Ca) encoding 60-kDa protein, has recently been discovered (C. Berry et al., Appl. Environ. Microbiol. 68:5082-5095, 2002). Cyt1Ca displays the structure two-domain fusion protein: N-terminal moiety resembles full-length...

In an attempt to endow Cyt1Ca with Cyt1Aa‐like antibacterial activity, both derived from Bacillus thuringiensis subsp. israelensis , two amino acids were replaced, E117V and N125A, so as raise the hydrophobicity of corresponding region, considered be membrane‐active motif. The clones obtained included multiple repeats VIEVLKSLLGIALA, head‐to‐tail polymerization primer, translated in frame Cyt1Ca. These versions caused instant arrest biomass growth decreased viability upon expression...

Gramineous plants protect their seeds from a variety of biotic stresses by producing toxic and deterrent secondary metabolites such as benzoxazinoids. It is unclear how the composition abundance these natural toxins has changed over course crop-plant domestication. To address this uncertainty, we characterized differences in metabolic levels benzoxazinoids derivatives, between four lines tetraploid wheat: wild emmer wheat (WEW), direct progenitor modern wheat; non-fragile domesticated (DEW),...

The microgene polymerization reaction (MPR) generates head-to-tail tandem repeats from homoduplexes (HDs). In MPR initiation, one HD putatively aligns two others in the proximity required to form a nucleation complex, thus allowing DNA polymerase skip intertemplate gap and generate an initial doublet (ID) prone repeat propagation. current investigation refines this stage by additional thermodynamic considerations elucidates fundamental mechanism underlying Four different types were designed...

One of the most important crops worldwide is wheat. Wheat domestication took place about 10,000 years ago. Not only that its wild progenitors have been discovered and phenotypically characterized, but their genomes were also sequenced compared to modern While comparative genomics essential track genes contribute improvement in crop yield, analyses functional biological end-products, such as metabolites, are still lacking. With advent rigorous mass-spectrometry technologies, it now possible...

Urethral infections caused by an emerging nongroupable (NG) urethrotropic clade of Neisseria meningitidis were first reported in the United States 2015 (the "U.S. NmNG urethritis clade"). Here, we evaluate for presence other urethral pathogens men with U.S. infection. We evaluated 129 urine specimens collected from at a sexual health clinic, including 33 patients culture-confirmed or suspected N. infection and 96 which nucleic acid amplification test detected gonorrhoeae, Chlamydia...

ABSTRACT dGTP starvation, a newly discovered phenomenon in which Escherichia coli cells are starved specifically for the DNA precursor dGTP, leads to impaired growth and, ultimately, cell death. Phenomenologically, it represents an example of nutritionally induced unbalanced growth: mass amplifies normally as dictated by nutritional status medium, but content is impaired. The other known such condition, thymineless death (TLD), involves starvation dTTP, has been found have important...

Togo has reported seasonal meningitis outbreaks caused by non-Neisseria meningitidis serogroup A (NmA) pathogens since the introduction of meningococcal conjugate vaccine (MACV, MenAfriVac) in 2014. From 2016 to 2017, NmW several outbreaks. In early 2019, a NmC outbreak was detected Savanes region and its investigation is described here. Under case-based surveillance, epidemiological clinical data, cerebrospinal fluid specimens were collected for every suspected case meningitis. Specimens...

A new gene, <i>cry11Bb2</i> from a field isolate of <i>Bacillus thuringiensis,</i> was cloned for expression in <i>Escherichia coli</i>. The encoded protein, with deduced molecular mass 89.5 kDa, exhibits 97 and 79% identities the overlap regions Cry11Bb1 <i>B. thuringiensis</i> ssp. <i>medellin</i> Cry11Ba1 <i>jegathesan</i>, respectively. It is however longer than by 42 amino acids its carboxy-terminus, which 32...