In Vivo and in Vitro Chromosome Replication in Bacillus subtilis

1973 ◽  
pp. 385-404 ◽  
Author(s):  
Noboru Sueoka ◽  
Tatsuo Matsushita ◽  
Seigou Ohi ◽  
Aideen O’Sullivan ◽  
Kalpana White
2009 ◽  
Vol 191 (6) ◽  
pp. 1749-1755 ◽  
Author(s):  
Jeffrey G. Gardner ◽  
Jorge C. Escalante-Semerena

ABSTRACT This report provides in vivo evidence for the posttranslational control of the acetyl coenzyme A (Ac-CoA) synthetase (AcsA) enzyme of Bacillus subtilis by the acuA and acuC gene products. In addition, both in vivo and in vitro data presented support the conclusion that the yhdZ gene of B. subtilis encodes a NAD+-dependent protein deacetylase homologous to the yeast Sir2 protein (also known as sirtuin). On the basis of this new information, a change in gene nomenclature, from yhdZ to srtN (for sirtuin), is proposed to reflect the activity associated with the YdhZ protein. In vivo control of B. subtilis AcsA function required the combined activities of AcuC and SrtN. Inactivation of acuC or srtN resulted in slower growth and cell yield under low-acetate conditions than those of the wild-type strain, and the acuC srtN strain grew under low-acetate conditions as poorly as the acsA strain. Our interpretation of the latter result was that both deacetylases (AcuC and SrtN) are needed to maintain AcsA as active (i.e., deacetylated) so the cell can grow with low concentrations of acetate. Growth of an acuA acuC srtN strain on acetate was improved over that of the acuA + acuC srtN strain, indicating that the AcuA acetyltransferase enzyme modifies (i.e., inactivates) AcsA in vivo, a result consistent with previously reported in vitro evidence that AcsA is a substrate of AcuA.


1981 ◽  
Vol 145 (2) ◽  
pp. 958-965 ◽  
Author(s):  
A H Ullah ◽  
G W Ordal
Keyword(s):  

2020 ◽  
Author(s):  
Erika Chacin ◽  
Priyanka Bansal ◽  
Karl-Uwe Reusswig ◽  
Luis M. Diaz-Santin ◽  
Pedro Ortega ◽  
...  

The replication of chromosomes during S phase is critical for cellular and organismal function. Replicative stress can result in genome instability, which is a major driver of cancer. Yet how chromatin is made accessible during eukaryotic DNA synthesis is poorly understood.Here, we report the identification of a novel class of chromatin remodeling enzyme, entirely distinct from classical SNF2-ATPase family remodelers. Yta7 is a AAA+-ATPase that assembles into ~ 1 MDa hexameric complexes capable of segregating histones from DNA. Yta7 chromatin segregase promotes chromosome replication both in vivo and in vitro. Biochemical reconstitution experiments using purified proteins revealed that Yta7’s enzymatic activity is regulated by S phase-forms of Cyclin-Dependent Kinase (S-CDK). S-CDK phosphorylation stimulates ATP hydrolysis by Yta7, promoting nucleosome disassembly and chromatin replication.Our results present a novel mechanism of how cells orchestrate chromatin dynamics in co-ordination with the cell cycle machinery to promote genome duplication during S phase.


1996 ◽  
Vol 43 (1) ◽  
pp. 107-114 ◽  
Author(s):  
J K Setlow ◽  
B C Setlow ◽  
P Setlow

The wild-type Bacillus subtilis spore protein, SspCwt, binds to DNA in vitro and in vivo and changes the conformation of DNA from B to A. Synthesis of the cloned SspCwt gene in Escherichia coli also causes large increases in mutation frequency. Binding of SspCwt to transforming DNA from Haemophilus influenzae made the DNA resistant to ultraviolet (UV) radiation. The mutant protein, SspCala, which does not bind DNA, did not change the UV resistance. The UV sensitivity of the DNA/SspCwt complex was not increased when the recipients of the DNA were defective in excision of pyrimidine dimers. These data indicate that the H. influenzae excision mechanism does not operate on the spore photoproduct formed by UV irradiation of the complex. Selection for the streptomycin- or erythromycin-resistance markers on the transforming DNA evidenced significant mutations at loci closely linked to these, but not at other loci. SspCwt apparently entered the cell attached to the transforming DNA, and caused mutations in adjacent loci. The amount of such mutations decreased when the transforming DNA was UV irradiated, because UV unlinks linked markers.


Author(s):  
Mihaela DUMITRU ◽  
Mihaela HĂBEANU ◽  
Cristina TABUC ◽  
Ștefana JURCOANE

This study aimed to evaluate some probiotic properties of Bacillus subtilis ATCC 6051a. The phenotypic profile, resistance to pH by simulated gastric juice (pH 2 and 3), bile salts by simulated intestinal fluid, survivability (%), heat and antibiotics tolerance were investigated. The strain is a Gram-positive, rod-shaped bacteria, arranged in short chains or in small irregular pairs with the ability to produce spores. Good viability at pH 2 and 3, with a survival of more than ≥80%, was found. In the presence of bile salts 0.3%, over 4 h, the strain exhibited a survival ≥85%. At 80°C, for 120 min., the strain showed good growth (9.04 log CFU/ml). Results were sensitive to most antibiotics, with a highly susceptible (between 16 – 25 mm) to erythromycin, clindamycin, amoxicillin, chloramphenicol, ciprofloxacin, amikacin and kanamycin. The strain was found to be sensitive to vancomycin, gentamicin, and tetracycline. The present research demonstrated that Bacillus subtilis ATCC 6051a can survive under gastrointestinal conditions, which involves them to future in vitro and in vivo probiotic studies.


2005 ◽  
Vol 73 (1) ◽  
pp. 31-36 ◽  
Author(s):  
Y. Douville ◽  
J.G. Boland

The influence and mechanisms of action of Bacillus subtilis on Colletotrichum trifolii, a causal agent of anthracnose of alfalfa (Medicago sativa), were studied in vivo and in vitro. In growth room conditions, a cell-free culture filtrate of B. subtilis significantly reduced disease incidence and severity on alfalfa seedlings from 56% to 16% and from 2.0 to 1.2, respectively. Treatment of seedlings with washed cell suspensions of B. subtilis had no influence on disease. Applications of crude filtrate on alfalfa leaflets inoculated with C. trifolii were associated with reduced germination of conidia, lysis of conidia, and reduced formation of appressoria. Under in vitro conditions, crude filtrate reduced germination of conidia, and induced lysis of conidia and the formation of inflated germ tubes on germinating conidia. An antibiotic of the iturin family, iturin D, was tentatively identified as the active compound responsible for the suppressive effect of B. subtilis on C. trifolii.


2021 ◽  
Vol 14 (2) ◽  
pp. 523-536
Author(s):  
Essam S. Soliman ◽  
Rania T. Hamad ◽  
Mona S. Abdallah

Background and Aim: Probiotics improve intestinal balance through bacterial antagonism and competitive exclusion. This study aimed to investigate the in vitro antimicrobial activity, as well as the in vivo preventive, immunological, productive, and histopathological modifications produced by probiotic Bacillus subtilis. Materials and Methods: The in vitro antimicrobial activities of B. subtilis (5×106 CFU/g; 0.5, 1.0*, 1.5, and 2.0 g/L) were tested against Escherichia coli O157: H7, Salmonella Typhimurium, Candida albicans, and Trichophyton mentagrophytes after exposure times of 0.25, 0.5, 1, and 2 h using minimal inhibitory concentration procedures. A total of 320 1-day-old female Ross broiler chickens were divided into five groups. Four out of the five groups were supplemented with 0.5, 1.0*, 1.5, and 2.0 g/L probiotic B. subtilis from the age of 1 day old. Supplemented 14-day-old broiler chickens were challenged with only E. coli O157: H7 (4.5×1012 CFU/mL) and S. Typhimurium (1.2×107 CFU/mL). A total of 2461 samples (256 microbial-probiotic mixtures, 315 sera, 315 duodenal swabs, and 1575 organs) were collected. Results: The in vitro results revealed highly significant (p<0.001) killing rates at all-time points in 2.0 g/L B. subtilis: 99.9%, 90.0%, 95.6%, and 98.8% against E. coli, S. Typhimurium, C. albicans, and T. mentagrophytes, respectively. Broilers supplemented with 1.5 and 2.0 g/L B. subtilis revealed highly significant increases (p<0.01) in body weights, weight gains, carcass weights, edible organs' weights, immune organs' weights, biochemical profile, and immunoglobulin concentrations, as well as highly significant declines (p<0.01) in total bacterial, Enterobacteriaceae, and Salmonella counts. Histopathological photomicrographs revealed pronounced improvements and near-normal pictures of the livers and hearts of broilers with lymphoid hyperplasia in the bursa of Fabricius, thymus, and spleen after supplementation with 2.0 g/L B. subtilis. Conclusion: The studies revealed that 1.5-2.0 g of probiotic B. subtilis at a concentration of 5×106 CFU/g/L water was able to improve performance, enhance immunity, and tissue architecture, and produce direct antimicrobial actions.


2006 ◽  
Vol 17 (2) ◽  
pp. 117-121 ◽  
Author(s):  
Ana Lúcia Campani Chassot ◽  
Maria Inês Pereira Poisl ◽  
Susana Maria Werner Samuel

The purpose of this study was to assess the antimicrobial efficacy of a peracetic acid-based disinfectant for decontamination of heat-polymerized, chemically activated and microwave-polymerized acrylic resins. Resin plates were contaminated in vivo upon intraoral use by 10 volunteers for 7 nights and slabs were contaminated in vitro by contact with Bacillus subtilis and Bacillus stearothermophilus. The contaminated acrylic resin specimens were immersed in a 0.2% peracetic acid-based disinfectant (Sterilife®; Lifemed) for 5 min or 10 min and placed in a BHI culture medium. After incubation at 37°C for 48 h, bacterial growth was assessed by analyzing turbidity of the medium. For all types of acrylic resin, no turbidity of the medium was observed for any of the resin specimens immersed in the peracetic acid-based disinfectant for either 5 or 10 min. On the other hand, the media with specimens that were not immersed in the disinfectant (control) showed turbidity in 100% of the cases, indicating the presence of microorganisms in both tested conditions. In conclusion, immersion for at least 5 min in a 0.2% peracetic acid-based disinfectant promoted high-level disinfection of heat-polymerized, chemically activated and microwave-polymerized acrylic resins contaminated with either human saliva or Bacillus subtilis or Bacillus stearothermophilus.


1998 ◽  
Vol 44 (8) ◽  
pp. 777-783 ◽  
Author(s):  
B M Sharga ◽  
G D Lyon

Antimicrobial substances were produced by Bacillus subtilis BS 107 in a defined medium and isolated from culture filtrate by precipitation at pH 2.5. Active fractions were extracted in ethyl acetate, acetone, and 80% ethanol and purified by thin-layer chromatography (TLC) on silica gel plates developed with an ethanol-water mixture (2:1, v/v). In each case, a band with a Rf of 0.75 formed an inhibitory zone when the TLC plates were placed in contact with agar seeded with test cultures of the Erwinia spp. The antibiotic was released into the culture medium during early stages of growth of Bacillus subtilis BS 107 but higher amounts were released in older cultures. The antibiotic was resistant to the action of nucleases, proteases, and lipase. It was stable when autoclaved twice for 35 min at 2 atm (1 atm = 101.325 kPa) in acidic, neutral, and alkaline solutions. It remained active over the pH range of 1-14 during 1 month of observation and exhibited no loss of antimicrobial activity when stored at 4°C for over 1 year. Bacillus subtilis BS 107 showed activity in vitro and in vivo against Erwinia carotovora subsp. atroseptica and Erwinia carotovora subsp. carotovora, the causal agents of potato blackleg and tuber soft rot. The application of an antagonist or its antibiotic to cut potato tissues prevented or reduced symptoms of the diseases. The antibiotic was active in vitro against a broad spectrum of bacterial and fungal species.Key words: antagonist, Bacillus subtilis BS 107, Erwinia carotovora, potato.


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