Gene/Protein
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Enzyme
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Target Concepts:
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Query: EC:3.1.25.1 (
deoxyribonuclease
)
1,471
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
The major conclusion from these studies is that variants of Haemophilus influenzae Rd which restrict and modify phage S2 are metastable and capable of giving rise to one another with high frequency. Nonrestrictive RdS cells segregate spontaneously to the restricting, modifying phenotype in about 5% of the progeny of a single clone. The restrictive cells derived from RdS revert to the nonrestrictive phenotype in 15 to 25% of the progeny of a single clone. These frequencies are not appreciably affected by treatment with acriflavine or ethidium bromide, compounds which affect plasmid stability, or by nitrosoguanidine, a powerful mutagen. The genetic locus for restriction and modification of bacteriophage S2 is found to have a chromosomal position between the biotin and proline loci. Restriction-modification of phage S2 has been shown to be a function of its deoxyribonucleic acid (DNA) in that transfection with S2 phage DNA or prophage DNA is subject to host restriction and modification. An enzyme preparation, which contains
endodeoxyribonuclease
but no appreciable exonuclease activity, from mutant
H. influenzae
com(-10) did not restrict phage S2.RdS DNA or prophage DNA transfecting activity, indicating that this
endodeoxyribonuclease
is not responsible for phage restriction. A new restriction enzyme isolated from
H. influenzae
Rd was found to be the major enzyme involved in the restriction of bacteriophage S2. The enzyme inactivated the transfecting activity of unmodified phage DNA but did not attack modified phage DNA. Unlike
endodeoxyribonuclease
R, this enzyme requires adenosine triphosphate and S-adenosylmethionine.
...
PMID:Restriction and modification of bacteriophage S2 in Haemophilus influenzae. 414 62
An enzyme similar to that described by Smith and Wilcox (15) for Haemophilus influenzae which attacks foreign deoxyribonucleic acid (DNA) but not its own has been isolated and purified from H. parainfluenzae. The enzyme degrades foreign DNA to limited sizes and can destroy the transforming activity of
H. influenzae
and Bacillus subtilis DNA. The enzyme can also destroy the biological activity of
H. influenzae
phage and prophage DNA. On the other hand, the
H. influenzae
endodeoxyribonuclease
can destroy the transforming activity of H. parainfluenzae DNA but not its own DNA. It also attacks B. subtilis DNA and its transforming activity.
...
PMID:Action of haemophilus endodeoxyribonuclease on biologically active deoxyribonucleic acid. 433 66
Hemophilus parainfluenzae
endodeoxyribonuclease
was used to degrade the DNA of
H. influenzae
and to follow the biological activity of 14 markers associated with this DNA. It was found that some
H. influenzae
markers were completely inactivated by
endodeoxyribonuclease
treatment, while others appeared to retain all or almost all of their original activity. The bulk of the
H. influenzae
DNA was reduced to double-stranded pieces of the order of 8 x 10(5) to 1 x 10(6) daltons. Velocity sedimentation of the DNA in sucrose gradients disclosed that markers that retained biological activity were present in DNA particles that were of the order of 1 x 10(6) daltons or larger, and indicated a close correlation between the size of the DNA fragment and the amount of biological activity retained. These data suggest that H. parainfluenzae
endodeoxyribonuclease
breaks DNA at specific sites. The nal(r) marker was shown to have twice as much biological activity after treatment with
endodeoxyribonuclease
when assayed at saturating DNA concentrations. In the linear portion of the DNA dose-response curve, the biological activity of this marker was reduced 3- to 10-fold compared to untreated DNA (in accord with the reduced size of its DNA). These data demonstrate a specific enrichment of the nal(r) marker by about 6- to 20-fold, and suggest a technique for the separation and purification of specific segments of DNA.
...
PMID:Separation of specific segments of transforming DNA after treatment with endodeoxyribonuclease. 463 56