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Biomedical subjects

J Vinograd

Publications and source records attributed to J Vinograd.

At least 37 records · Page 2Linked to original sources
PubMed1973

The presence of ribonucleotides in mature closed-circular mitochondrial DNA.

Mitochondrial DNA from mouse and HeLa cells is nicked by alkali and chick-embryo ribonuclease H. It has been concluded that ribonucleotides are present in the closed-circular duplex. A comparative study of the scission rates at pH 13.0 of RNA and of mitochondrial DNAs indicates that there are no more than 10 ribonucleotides in the major fraction of the mitochondrial DNAs.

Alkaline Phosphatase↗
PubMed1972

Sequence heterogeneity in closed simian virus 40 deoxyribonucleic acid.

The heteroduplex molecules formed by self-annealing of denatured, singly nicked simian virus 40 (SV40) deoxyribonucleic acid (DNA) prepared from closed viral DNA were examined by formamide-protein film electron microscopy to test the DNA for sequence homogeneity. Sequence inhomogeneity appears in the heteroduplexes as single-strand loops. These result from sequence deletion or from sequence substitution, if regions greater than 50 nucleotides are involved. The undenatured DNA from viruses passaged twice at multiplicities of infection much less than 1 plaque-forming unit (PFU) per cell appeared to be homogeneous in size. The heteroduplexes formed by this DNA indicated that approximately 2% of the molecules carried deletions, but that substitutions were below the level of detection. In contrast, undenatured DNA from viruses grown by passaging undiluted lysates seven times or by infection with stock virus at a multiplicity of infection of 5 PFU per cell contained a large frequency of molecules shorter than the full length. The heteroduplex samples indicated that 12 and 7% of the undenatured material contained base substitutions, and 13 and 11% contained deletions. The deletions and substitutions appear to occur in separate molecules. Length measurements on heteroduplexes displaying the loop characteristic of substitutions have established that these molecules are from true sequence substitutions, and not from adjacent or overlapping deletions. More than 80% of the molecules carrying substitutions are shorter than the native SV40 length. On the average, the substituted sequence is about 20% of the length of SV40, but it replaces a sequence about 30% of the native length. The substituted sequences may be host cell nuclear DNA, possibly arising from integration of SV40 into the chromosome followed by excision of the SV40 DNA together with chromosomal DNA.

Animals↗
PubMed1971

Superhelix density heterogeneity of intracellular simian virus 40 deoxyribonucleic acid.

Covalently closed intracellular and viral simian virus 40 (SV40) deoxyribonucleic acid (DNA) were separately isolated from infected African green monkey cells (BSC-1) grown in culture. The two DNA species form overlapping bands centered at different positions in a propidium di-iodide-cesium chloride (PDI-CsCl) buoyant density gradient capable of separating closed DNA species with different superhelix densities. When the dense side of a (32)P-labeled intracellular DNA band was mixed with the light side of a (3)H-labeled intracellular DNA band and again centrifuged in a PDI-CsCl density gradient, two overlapping bands formed with modes displaced from each other. Similar band-splitting experiments performed with viral DNA always gave superimposable bands. The foregoing experiments demonstrate that the intracellular DNA is heterogeneous in superhelix density, whereas, by the same criteria, the viral DNA is homogeneous. The mean superhelix density of the intracellular closed DNA is approximately three-fourths as large as the superhelix density of the viral DNA. These results rule out the possibility that closed SV40 DNA is drawn randomly from the intracellular pool and assembled without a further nicking-closing step into virions. When the cells were grown and infected in the presence of ethidium bromide (EB), the intracellular closed DNA was found to be homogeneous in superhelix density and to have the same superhelix density as the viral DNA which, in turn, was unaffected by the presence of the drug. The foregoing results were explained by postulating that the intracellular DNA is formed with a homogeneous superhelix density and becomes heterogeneous in the absence of EB as a result of a nicking-closing cycle that occurs in a spacially or temporally heterogeneous environment. The drug EB would inhibit this action by inhibiting the nicking enzyme(s).

Cell Line↗