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Query: EC:3.1.21.1 (
DNase
)
7,655
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
By a direct assay approach, mutants of Haemophilus influenzae Rd that are deficient in adenosine 5'-triphosphate-dependent
deoxyribonuclease
activity (add-) were isolated and characterized. A large proportion (50 to 90%) of the cells in cultures of these mutants failed to produce visible colonies when plated. An extensive analysis of the recombination proficiency of these strains revealed that the transformation frequency (transformants per competent cell) in the mutants was similar to that found in the wild type, but that the transformation efficiency (transformants per microgram of irreversibly bound deoxyribonucleic acid [DNA]) was reduced approximately fourfold. Sensitivities of the mutants to gamma rays, ultraviolet radiation, and methyl methane sulfonate were only slightly greater than wild-type levels. The rate of degradation of host DNA after ultraviolet irradiation was significantly reduced in the mutants. It is suggested that the adenosine 5'-triphosphate-dependent
deoxyribonuclease
in
H. influenzae
plays a nonessential role in DNA recombination and repair.
...
PMID:Isolation and characterization of mutants of Haemophilus influenzae deficient in an adenosine 5'-triphosphate-dependent deoxyribonuclease activity. 16 69
During bacteriophage studies on Haemophilus influenzer, it was observed that encapsulated type b and unencapsulated Rb strains released a bactericidal substance acitve against types a, c, d, e, and f
H. influenzae
, non-typable
H. influenzae
strains, other Haemophilus species, and certain members of the Enterobacteriaceae. The bactericidal activity was assayed by a plaque test utilizing an Rd strain as an indicator lawn and was also demonstrated in mixed broth cultures of a producer strain and an indicator strain. Immediately lysis of sensitive bacteria by the factor was not evident. The factor is sensitive to trypsin but resistant to
deoxyribonuclease
, treatment with 2-mercaptoethanol, lipase, alpha-amylase, and heating in a 100 degrees C water bath for 20 min. The activity is not dependent upon increased Ca2+ or Mg2+ concentration as is necessary for HP1C1 and S2 phage propagation. The bactericidal factor is not pelleted by high-speed centrifugation at 150,000 X g for 6 h. Treatment with ultraviolet light or mitomycin C does not result in observable phage, phage-like particles, or increased bactericidal activity. T-HE BACTERICIDAL FACTOR IS NOT A TYPICAL SMALL MOLECULAR WEIGHT "COLICIN-LIKE" BACTERiocin in that it is not inducible, has a wider range of activity, and does not kill by "single-hit" kinetics. On preliminary characterization, it is a thermostable protein toxic to certain bacterial strains.
...
PMID:Bactericidal substance produced by Haemophilus influenzae b. 108 28
A transformation-deficient strain of Haemophilus influenzae, lacking adenosine 5'-triphosphate-dependent
deoxyribonuclease
activity, was isolated by selection for sensitivity to mitomycin. The mutant, designated JK57, possibily showed a moderate sensitivity to ultraviolet (UV) irradiation and treatment with methyl methane sulfonate. Contrary to the wild type, the mutant degraded chromosomal deoxyribonucleic acid (DNA) to some extent. However, after UV irradiation to the mutant degraded considerably less DNA than the wild type and the TD24 mutant of
H. influenzae
, the latter being equivalent to a recA mutant of Escherichia coli. A TD2457 double mutant, constructed by transferring the TD24 mutation into the JK57 strain, was as sensitive to deleterious agents and as deficient in transformation as the TD24 single mutant; in the double mutant, however, after UV irradiation chromosomal DNA was degraded to the same extent as in the JK57 mutant. The number of transformants per unit of radioactive donor DNA taken up by JK57 recipient cells was approximately 10-fold smaller than in the wild type. Presynaptically, the fate of donor DNA in the adenosine 5'-triphosphate-dependent
deoxyribonuclease
-deficient mutants was not different from that in the wild type. In contrast to TD24 and the TD2457 double mutant, in the JK57 mutant, recombinant-type activities (molecules carrying both the donor and recipient markers) were formed almost as well as in the wild type. After integration into the JK57 recipient genome, the rate of replication of the donor marker was equal to that of the recipient marker during a number of generations, which suggests that the donor DNA is normally integrated into the JK57 chromosome. It is suggested that transformed JK57 cells pass with a high frequency into a type of cells that can replicate their chromosomes many times but have lost the ability to form visible colonies after plating.
...
PMID:Effect of adenosine 5'-triphosphate-dependent deoxyribonuclease deficiency on properties and transformation of Haemophilus influenzae strains. 108 3
Relationship between the normal throat flora and pathogenic bacteria recovered from the throat in 139 children with upper respiratory tract infections in winter were studied using quantitative analyses. Pathogenic bacteria examined include S. pyogenes,
H. influenzae
, S. aureus, and S. pneumoniae, and the normal floras include alpha-streptococci, gamma-streptococci, Neisseria species, and Micrococci. Children with S. pyogenes in their throats (S. pyogenes group) were examined with anti-streptococcal antibodies such as anti-streptolysin O, anti-streptokinase, and anti-
deoxyribonuclease
B. Eighty seven pathogenic bacteria were recovered from 72 children (51.8%) out of 139. S. pyogenes and S. pneumoniae groups showed significantly lower alpha-streptococci and gamma-streptococci in incidence of appearance when compared with children with the no pathogenic bacteria in their throats (no bacteria group).
H. influenzae
group showed significantly lower gamma-streptococci and higher Neisseria sp. in incidence of appearance compared with the no bacteria group. Positive cases for anti-streptococcal antibodies showed a significantly lower alpha-streptococci in number compared with negative cases for antibodies and the no bacteria group, and a significantly lower gamma-streptococci in incidence of appearance compared with the no bacteria group. These data suggest that the normal throat flora may have a role in prevention of colonization by the pathogenic bacteria in vivo, as were shown in vitro by many authors, and that the quantitative analysis of the normal flora is useful because this methodology might reveal whether the bacteria recovered from the throat show the pathogenicity.
...
PMID:[Quantitative analyses of the normal throat flora of children with upper respiratory tract infections]. 140 17
We previously demonstrated that pneumococcal extracts contain a highly specific inhibitor of human neutrophil elastase (HNE). We now show that the active inhibitor in these extracts is a high-molecular-weight, heat-stable substance that appears to be RNA, since inhibitory activity of pneumococcal extracts is decreased by incubation with ribonuclease but not by incubation with
deoxyribonuclease
or proteinase K. Moreover, metabolically labeled ([3H]uridine) pneumococcal RNA, isolated by phenol extraction followed by ethanol precipitation, strongly inhibits HNE. Pneumococcal capsular polysaccharide, although polyanionic, is only weakly inhibitory toward HNE and is not a major source of elastase-inhibitory activity in pneumococcal extracts. On the other hand, the capsule of Haemophilus influenzae type b contains polyribosylribitol phosphate. This highly charged polyanion possesses HNE-inhibitory activity, but only under special circumstances to be discussed below. Pneumococci (type I, type II smooth, type II rough) and
H. influenzae
(type b) all release HNE-inhibitory activity into their culture medium during growth. By contrast, Klebsiella pneumoniae and Staphylococcus aureus release little (if any) stable HNE-inhibitory activity during growth. We propose that some bacterial pneumonias may spare host tissue because polyanions released by the invading microorganisms (e.g. RNA from autolysing pneumococci) inhibit elastase released from inflammatory neutrophils and thereby modulate accompanying tissue proteolysis. Pneumonias caused by microorganisms that do not release stable polyanionic inhibitors of HNE (e.g., Staphylococcus and Klebsiella) may be correspondingly more injurious to the lung.
...
PMID:Inhibition of human neutrophil elastase by bacterial polyanions. 244 47
Transformation pathways in two closely related bacterial species, Haemophilus parainfluenzae and Haemophilus influenzae, were studied. Both organisms rapidly take up transforming DNA within minutes into specialized membranous structures on the cell surface (transformasomes). DNA within transformasomes is in a protected state, inaccessible to external
DNase
or internal restriction and modification enzymes. However, the subsequent processing of donor DNA differs in these two organisms. In
H. influenzae
, linear DNA immediately undergoes degradation from one end at a constant rate, leaving a lower-molecular-weight intermediate in the transformasome. The end undergoing degradation is searching for homologous regions of the chromosome. Once pairing is initiated, the remaining lower-molecular-weight DNA exits from the transformasome, and a single strand undergoes efficient integration. In contrast, in H. parainfluenzae little degradation of donor DNA is observed, with the majority remaining intact within the transformasomes after 1 h. Thus, whereas only 10% of donor DNA molecules leave the protected state after 1 h, portions of each molecule appear to become quantitatively integrated.
...
PMID:Comparison of transformation mechanisms of Haemophilus parainfluenzae and Haemophilus influenzae. 298 12
In 82 patients with acute tonsillitis studied, beta-hemolytic group A streptococci were isolated from 30 (37%), and group C or G streptococci from 12 (15%). In the 40 patients with non-streptococcal tonsillitis there was a significantly higher isolation rate of pneumococci,
H. influenzae
and/or B. catarrhalis, as compared with those with beta-hemolytic streptococci. Patients were classified regarding clinical status according to standardized criteria as severe, moderate, or mild. The patients with group A streptococcal tonsillitis were significantly more often classified clinically as 'severe' and had significantly shorter duration of symptoms before seeking medical care, as compared with those with non-streptococcal tonsillitis. Significant increases in white blood cell count and in anti-
DNase
B were found in the patients with group A streptococcal tonsillitis, whereas their antistreptolysin O levels did not increase significantly. C-reactive protein concentrations were consistently higher in the patients with group A streptococcal tonsillitis. No evidence of polyclonal beta-lymphocyte stimulation was found when measuring antibodies against pneumococci and group B streptococci. The findings show clinical and simple laboratory tests to be useful aids in distinguishing group A streptococcal tonsillitis from non-streptococcal tonsillitis, and that other bacteria may be involved in non-streptococcal tonsillitis.
...
PMID:Clinical and laboratory findings in patients with acute tonsillitis. 331 20
Local bending propensity and curvature of DNA can be characterized using a vector description of DNA bendability, based on a set of parameters derived from
deoxyribonuclease I
(DNase I) cleavage experiments. Two characteristics-arithmetic and vector averages of bendability-were successfully used to predict experimentally known bendable, rigid and curved segments in DNA. A characteristic distribution of bendability is conserved in evolutionarily related kinetoplast sequences. An analysis of the M. genitalium and
H. influenzae
genomes as well as fragments of human and yeast genomes shows, on the other hand, that highly curved segments--similar to artificially designed curved oligonucleotides--are extremely rare in natural DNA.
...
PMID:Distribution of bending propensity in DNA sequences. 880 40
INTRODUCTION: Between 20% and 50% of middle ear effusions in otitis media with effusion test positive for bacteria, i.e.
H. influenzae
, M. catarrhalis and S. pneumoniae. In one study 48% of effusions that tested negative by culture wre positive by polymerase chain reaction (PCR).1 This has been advanced as evidence for the presence of bacterial biofilms, as agents leading to the persistence of glue ear. There is, however, the possibility that the DNA detected is the fossilized remains of bacteria from previously cleared infections. If this is the case, then the effusioin must in some way be protecting the DNA from breakdown by DNases. METHODS: Here we demonstrate, using a viscosity assay, that middle ear effusions taken from children during myringotomy inhibit the breakdown of DNA in a concentration-dependent manner. Middle ear effusion homogenates 0.5-3.0 ml (1 : 10 Vol. vol/effusion: PBS) were incubated with 3 ml of DNA (0.5 mg/ml in PBS) plus 0.2 ml of
DNase
1 (500 Kunitz) at 37 degrees C for 24 h. Viscosity measurements were taken at regular intervals and the changes in viscosity expressed as a percentage of time 0.
DNase
activity was inhibited by 1.5 ml and 3.0 ml of effusion 48% and 91%, respectively, after 30 min incubation. CONCLUSION: This study demonstrated that effusions have the ability to inhibit nuclease activity, and the reported presence of non-culturable bacteria based on DNA detection by PCR could still represent fossilized remains and not viable bacteria. The same could also be true of recent reports of bacterial mRNA.2
...
PMID:Does the bacterial DNA found in middle ear effusions come from viable bacteria? 1112 7