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At least 703 records · Page 39Linked to original sources
PubMed1989

Locations of nucleosomes on the regulatory region of simian virus 40 chromatin.

We have asked where the nucleosomes are located with respect to the replication origin and regulatory region of simian virus 40 DNA, what would be the possible functional consequences of the identified locations, and to what extent these locations correlate with the current views on mechanisms involved in establishing nucleosome-free regions in chromatin. To identify the precise location of nucleosomes, we have shot-gun cloned and sequenced nucleosomal DNA obtained from micrococcal nuclease digestion of wt776 chromatin prepared late in infection. Our results indicate that nucleosomes do not occupy unique positions over the replication origin or the elements involved in transcriptional control. However, it appears that the nucleosome distribution is not random, since several nucleosomes are represented by two or more independently generated clones. Two nearly identical cloned fragments map over the replication origin; five include 1.5 copies of the 72 base-pair enhancer sequences; and eight map to a region that spans a DNA bending locus and the major transcription initiation site of the late genes. The complex nucleosome distribution pattern observed in our direct analysis suggests that disparate nucleosome-free regions may be involved in controlling replication, and selective expression of the viral early or late genes.

Cell Line↗
PubMed1986

Chromatin structure in scrapie and Alzheimer's disease.

Scrapie affected brains exhibit a number of pathological features in common with the human neurodegenerative condition, Alzheimer's disease. The present report describes studies on chromatin structure seen in these two disease processes. Chromatin associated proteins influence transcriptional activity of DNA through an effect upon chromatin structure. We examined chromatin structure by: measuring the capacity of the enzyme micrococcal nuclease to release mono- and dinucleosomes from isolated nuclei and measuring DNA-histone interactions by examining the effect of ambient tonicity upon the release of chromatin proteins. In two strains of mice infected with two strains of scrapie agent there was reduced accessibility to micrococcal nuclease and an increased content on dinucleosomes of the histone H1 and H1(0) types. These changes precede clinical signs of scrapie and resemble those found in the human conditions of Alzheimer's and Pick's disease. Scrapie mouse brain differs from Alzheimer brain in that scrapie does not alter histone-DNA interactions as monitored by ionically induced histone release from chromatin. Despite similarities, the scrapie agent appears to operate upon different molecular mechanisms than those found in Alzheimer's disease.

Alzheimer Disease↗
PubMed1978

Cell-free systems for in vitro translation of herpes simplex virus messenger RNA.

A cell-free system active in translation of HSV mRNA was obtained by fractionation of reticulocyte lysates. Endogenous protein synthesis was further reduced by preincubation with micrococcal nuclease, although this treatment did not affect the translation of infected (or uninfected) BHK cell RNA. Polypeptides synthesized by the fractionated reticulocyte cell-free system were very similar to those produced by unfractionated reticulocyte lysates.

Cell Fractionation↗
PubMed1986

Chromosomal organization of the herpes simplex virus genome during acute infection of the mouse central nervous system.

After corneal inoculation, herpes simplex virus type 1 replicates in the mouse eye, trigeminal ganglia, and brainstem, producing first an acute and then a latent infection. Previous work from this laboratory focused on the structure of the viral DNA in this system. We have now examined the structure of the viral genome at the chromosome level by using micrococcal nuclease digestion. Studies with disaggregated cell preparations made from the brainstems of acutely infected mice show that the majority of the viral DNA is in a nonnucleosomal form; however, a nucleosomelike fraction was also consistently detected. A similar result was obtained for viral DNA in herpes simplex virus type 1-infected C1300 (clone NA) neuroblastoma cells (a neuronal cell line).

Acute Disease↗
PubMedDate not supplied

[Development of gentamicin-resistant Small Colony Variants of S. aureus after implantation of gentamicin chains in osteomyelitis as a possible cause of recurrence].

AIM: Recently, S. aureus small colony variants (SCVs) were reported to persist within cultured endothelial cells and to cause persistent and antibiotic resistant infections in humans. Because gentamicin can very reproducably select for electron transport deficient SCVs as shown in earlier in vitro experiments, we searched for SCVs in a patient with chronic osteomyelitis, who received gentamicin beads. METHOD: Special culture and identification procedures for determination of SCVs were used, including testing of the S. aureus specific nuc gene, pulsed field gel electrophoresis (PFGE) as a typing method and characterization of the auxotrophism of the SCVs. RESULTS: In a case of a 34-year-old patient with chronic osteomyelitis who had previously been treated with gentamicin beads, menadione auxotrophic S. aureus SCVs as well as wild type S. aureus were recovered in multiple bone specimen. All different colony types isolated from simultaneous or from sequential specimen were shown to be clonal by PFGE. CONCLUSION: Recovery of S. aureus SCVs from a patient treated with gentamicin beads suggests that the slow release of gentamicin into the local environment may be an efficient way to select for and/or induce SCVs. These data should alert physicians to also consider SCVs when a treatment failure occurs in a patient that has received gentamicin beads.

Adult↗
PubMed1983

Serology of Staphylococcus aureus infections using multiple antigens and serial serum samples.

The detection of antibody to S aureus in human serum can aid in the management of staphylococcal diseases [1]. RIAs and ELISAs can detect low levels of antibody and demonstrate increased antibody production during serious staphylococcal infections [2,3]. We compared four S aureus constituents--cell-wall peptidoglycan and teichoic acid, and extracellular alpha-toxin and nuclease--as antigens in a sensitive ELISA. The value of testing more than a single serum sample was also determined. Elevated IgG antibody to peptidoglycan, present in one or more serum samples of 13 (50%) of 26 patients with complicated bacteremia, was found to be the most sensitive test. All 26 patients had a significant IgG antibody response to peptidoglycan. Three (27%) of 11 patients with uncomplicated bacteremia had elevated levels of antibody to peptidoglycan in their serum, and seven (64%) showed a significant change in titer when serial serum samples were tested. Maximum detection rates for the other antigens in complicated and uncomplicated bacteremia were, respectively, 62% and 37% for teichoic acid, 38% and 37% for nuclease, and 54% and 13% for alpha-toxin. In single serum samples, the detection rate for all four antigens marginally improved the results, with detection rates of 62% and 36% for complicated and uncomplicated bacteremia, respectively. Cross-reactive antibody to peptidoglycan but not to the other three antigens was present in six (75%) of eight patients with long-standing subacute bacterial endocarditis due to either viridans streptococci or Staphylococcus epidermidis (data not shown).(ABSTRACT TRUNCATED AT 250 WORDS)

Antibodies, Bacterial↗
PubMed1989

Identification of factors specific for transcription of the late class of vaccinia virus genes.

Cytoplasmic extracts made from HeLa cells that have been harvested late after infection with vaccinia virus are capable of specifically transcribing templates containing vaccinia virus late-gene promoters. We applied such an extract to a phosphocellulose column and eluted the proteins with a series of buffers containing successively higher concentrations of NaCl. None of three column fractions alone was capable of specific transcription of a late-gene template. However, specific transcriptase activity could be reconstituted by mixing column fractions, with maximal activity seen when all three fractions were present. The activities present in all fractions were heat labile, resistant to micrococcal nuclease, and present only in extracts from vaccinia virus-infected cells. A quantitative complementation assay was used to further purify one factor, named VLTF-1, over subsequent columns of DEAE-cellulose and hydroxylapatite. VLTF-1 was separated from endogenous RNA polymerase, was a late-promoter-specific transcription factor, and had a sedimentation rate consistent with an apparent Mr of 45,000. The RNA polymerase-containing fraction was not only necessary for transcription with a late-promoter template but alone was capable of specifically transcribing a vaccinia virus early-gene promoter. A further difference between early and late gene transcription in this system was in the ability of the ATP analog beta-8-imidoadenosine-5'-triphosphate (AMP-PNP) to substitute for ATP in supporting specific transcription of only the late-promoter template. The system reconstituted from the various fractions retained the ability to produce the novel poly(A) sequence found on the 5' end of vaccinia virus late messages.

Adenosine Triphosphate↗