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  • 1
    Electronic Resource
    Electronic Resource
    Mammalian .alpha.-polymerase: cloning of partial complementary DNA and immunobinding of catalytic subunit in crude homogenate protein blots (1987)
    s.l. : American Chemical Society
    Biochemistry 26 (1987), S. 956-963 
    Details
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1021/bi00377a041
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    Paper (German National Licenses)
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  • 2
    Electronic Resource
    Electronic Resource
    Interaction of simian virus 40 large T-antigen with cellular DNA polymerase α: studies with various T-antigen mutants of SV40 (1991)
    Springer
    Archives of virology 118 (1991), S. 113-125 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Simian virus 40 (SV40) large T antigen binds to host cell DNA polymerase α and the T-antigen-DNA polymerase α complex is implicated in the initiation of viral DNA replication. We have examined various SV40 T-antigen mutants to test the correspondence between viral DNA replication and T-antigen-DNA polymerase α complex formation. The various SV40 T-antigen mutants were used to either infect or transfect African green monkey kidney cell line CV-1, and at different time intervals we measured the production of T-antigen and host cell DNA polymerase α by radioimmunoassay, complex formation by a sandwich radioimmunoassay and the amount of viral DNA synthesis by dot-blot hybridization analysis. There was a good correlation between complex formation and viral DNA synthesis in lytic mutants of SV40. Poor complex formation and correspondingly lower DNA synthesis were observed in the non-viable mutants of SV40, even though significant amounts of T-antigen and DNA polymerase α were present. Our results substantiate the earlier findings of T-antigen-DNA polymerase α complex formation and establish the need for formation of this complex in promoting viral DNA synthesis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01311307
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  • 3
    Electronic Resource
    Electronic Resource
    Chinese hamster ovary cell mutants resistant to DNA polymerase inhibitors (1985)
    Springer
    Molecular genetics and genomics 200 (1985), S. 393-400 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Isolation and characterization of Chinese hamster ovary cell mutants resistant to different DNA polymerase ase inhibitors (aphidicolin, ara-A and ara-C) have been described. A particular mutant (JK3-1-2A) characterized in detail was found to grow and synthesize DNA in medium containing an amount of aphidicolin tenfold greater than that which completely inhibited the growth and the DNA synthesis of the wild-type cells. An almost twofold increase in the specific activity of the DNA polymerase α was seen in this mutant. The mutant DNA polymerase showed altered aphidicolin inhibition kinetics of dCMP incorporation; the apparent K m for dCTP and the apparent K i for aphidicolin were increased in the mutant. These alterations in the kinetic parameters were, however, abolished upon further purification of the enzyme. Ara-CTP was found to act as a competitive inhibitor of the dCMP incorporation by both the wild type and mutant enzymes. In contrast, the effect of aphidicolin on dCMP incorporation was either competitive (wild-type enzymes) or noncompetitive (mutant enzyme). The data presented showed that the sites of action for aphidicolin and ara-CTP were distinct; likewise the dCTP binding site appeared to be separate from other dNTP(s) binding sites. The drug resistance of the mutant was inherited as a dominant trait.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425722
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  • 4
    Electronic Resource
    Electronic Resource
    Chinese hamster ovary cell mutants resistant to DNA polymerase inhibitors (1985)
    Springer
    Molecular genetics and genomics 200 (1985), S. 401-406 
    Details
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Genetic characterizations of the Chinese hamster ovary cell mutants resistant to the DNA polymerase inhibitors (aphidicolin, ara-A and ara-C) have been described. Resistance to all three inhibitors showed dominance among the progeny of somatic cell crosses between the wild type and mutant parents. Analysis of the segregation of the drug-resistant character among 566 hybrid progeny from somatic crosses between the wild type (aph s, ara-A s, and ara-C s) and the triple mutants (aph r, ara-A r, ara-C r) showed the involvement of at least three unlinked genes in controlling the expression of the resistance to different DNA polymerase inhibitors. The mutant (aph r) DNA was used to transfect aphidicolin resistance to recipient mouse Ltk- cells either directly or in combination with the plasmid pTK2 DNA. The aphidicolin resistance of the transfected cells was found to be a stable phenotype and could be used in multiple rounds of transfection, indicating the chromosomal integration of the transfecting gene.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00425723
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    Paper (German National Licenses)
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