Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.776367
Title: Enzymes of DNA synthesis and degradation in cells infected with herpes simplex virus
Author: Paton, Robert D.
Awarding Body: University of Glasgow
Current Institution: University of Glasgow
Date of Award: 1972
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Abstract:
The main aim of the project was to purify the virus-induced DNA polymerase from herpes simplex virus-infected baby hamster kidney cells grown in culture. If this could not be carried to homogeneity, then the aim would be at lea.st to separate host DNA polymerase and DNase from the enzyme in order to clarify the relationship of the induced DNA polymerase and DNase and eliminate the interfering effects of the other enzymes. In. the course of the present purification studies involving column chromatography on DEAE-cellulose, hydroxylapatite, Sephhadex G-150, and phosphocellulose, and fractionations using sucrose gradient sedimentation and polyacrylamaide gel electrophoresis, evidence was obtained that at least part of the virus-induced DNase activity could be separated from DNA polymerase especially on hydroxylapatite columns. The question arose whether there were one or two new virus-induced DNA exomuclease(s) because, while one of the peaks of DNase from hydroxylapatite was izolymerase-free the other was closely associated with polymerase and remained so throughout several purification steps. The two virus-induced exonuclease peaks wore compared using several criteria including: sedimentation coefficients, substrate specificities, heat stability and rechromatography. The results showed that the enzymes were in the main similar by these criteria although differences detected by the last two criteria may be significant. On the whole these and other characterisation studies coupled with consideration of earlier work suggested that there was probably only one virus-induced DNA exonuclease, and that this was distinct from the DNA polymerase purified protein. Nevertheless, there is also substantial evidence in favour of there being two distinct virus-induced exonucleases, one polymerase-free, the other polymerase-associated. A further possibility that the polymerase-free exonuclease is a breakdown product of a single virus-induced DNA polymerase-DNA exonuclease protein has not been eliminated completely by studies carried out with a protease inhibitor. The final answer rests upon further purification studies.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.776367  DOI: Not available
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