Please use this identifier to cite or link to this item: http://dlib.scu.ac.ir/handle/2027.42/24900
Title: DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs
Authors: School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A
Publisher: Elsevier
Description: Nuclei isolated from herpes simplex virus (HSV) type 2-infected KB cells were examined for their capacity to serve as an in situ source of herpes DNA polymerase. In contrast to purified enzymes with added template, approx. 80% of the DNA synthesized in isolated nuclei was viral. The average size of DNA fragments labeled in vitro was 3.2[middle dot]106 Da. Based on an increase in DNA density when nuclei were incubated in the presence of BrdUTP rather than dTTP, 16% of the nucleotides were added during the in vitro reaction. Sucrose gradient analysis of DNA polymerase activity in extracts of isolated nuclei demonstrated the nearly exclusive presence of herpes DNA polymerase. Km concentrations for the four dNTPs were from 0.14 to 0.55 [mu]M. DNA synthesis was inhibited competitively by the 5'-triphosphates of ara-A and ara-C (Ki = 0.03 and 0.22 [mu]M, respectively) but not by the 5'-triphosphate of dideoxythymidine. aATP also served as a substrate (Km = 0.014 [mu]M) for the reaction. We conclude that nuclei from HSV-infected cells have significant advantages for the detailed study of inhibitors of herpesvirus replication.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24900
More Information: Barnett, Jimmy W., Reinke, C. Michael, Turk, Steven R., Drach, John C. (1984/02/24)."DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs." Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 781(1-2): 130-142. <http://hdl.handle.net/2027.42/24900>
http://www.sciencedirect.com/science/article/B6T1V-47S5YFF-H9/2/1028d1c25c4c921d5dc98e29cb8dbbe1
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6320890&dopt=citation
http://hdl.handle.net/2027.42/24900
6320890
http://dx.doi.org/10.1016/0167-4781(84)90131-3
Biochimica et Biophysica Acta
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Files in This Item:
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Title: DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs
Authors: School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A
Publisher: Elsevier
Description: Nuclei isolated from herpes simplex virus (HSV) type 2-infected KB cells were examined for their capacity to serve as an in situ source of herpes DNA polymerase. In contrast to purified enzymes with added template, approx. 80% of the DNA synthesized in isolated nuclei was viral. The average size of DNA fragments labeled in vitro was 3.2[middle dot]106 Da. Based on an increase in DNA density when nuclei were incubated in the presence of BrdUTP rather than dTTP, 16% of the nucleotides were added during the in vitro reaction. Sucrose gradient analysis of DNA polymerase activity in extracts of isolated nuclei demonstrated the nearly exclusive presence of herpes DNA polymerase. Km concentrations for the four dNTPs were from 0.14 to 0.55 [mu]M. DNA synthesis was inhibited competitively by the 5'-triphosphates of ara-A and ara-C (Ki = 0.03 and 0.22 [mu]M, respectively) but not by the 5'-triphosphate of dideoxythymidine. aATP also served as a substrate (Km = 0.014 [mu]M) for the reaction. We conclude that nuclei from HSV-infected cells have significant advantages for the detailed study of inhibitors of herpesvirus replication.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24900
More Information: Barnett, Jimmy W., Reinke, C. Michael, Turk, Steven R., Drach, John C. (1984/02/24)."DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs." Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 781(1-2): 130-142. <http://hdl.handle.net/2027.42/24900>
http://www.sciencedirect.com/science/article/B6T1V-47S5YFF-H9/2/1028d1c25c4c921d5dc98e29cb8dbbe1
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6320890&dopt=citation
http://hdl.handle.net/2027.42/24900
6320890
http://dx.doi.org/10.1016/0167-4781(84)90131-3
Biochimica et Biophysica Acta
Appears in Collections:Research Collections

Files in This Item:
Click on the URI links for accessing contents.
Title: DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs
Authors: School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A;School of Dentistry, University of Michigan, Ann Arbor, MI 48109, U.S.A
Publisher: Elsevier
Description: Nuclei isolated from herpes simplex virus (HSV) type 2-infected KB cells were examined for their capacity to serve as an in situ source of herpes DNA polymerase. In contrast to purified enzymes with added template, approx. 80% of the DNA synthesized in isolated nuclei was viral. The average size of DNA fragments labeled in vitro was 3.2[middle dot]106 Da. Based on an increase in DNA density when nuclei were incubated in the presence of BrdUTP rather than dTTP, 16% of the nucleotides were added during the in vitro reaction. Sucrose gradient analysis of DNA polymerase activity in extracts of isolated nuclei demonstrated the nearly exclusive presence of herpes DNA polymerase. Km concentrations for the four dNTPs were from 0.14 to 0.55 [mu]M. DNA synthesis was inhibited competitively by the 5'-triphosphates of ara-A and ara-C (Ki = 0.03 and 0.22 [mu]M, respectively) but not by the 5'-triphosphate of dideoxythymidine. aATP also served as a substrate (Km = 0.014 [mu]M) for the reaction. We conclude that nuclei from HSV-infected cells have significant advantages for the detailed study of inhibitors of herpesvirus replication.
URI: https://deepblue.lib.umich.edu/handle/2027.42/24900
More Information: Barnett, Jimmy W., Reinke, C. Michael, Turk, Steven R., Drach, John C. (1984/02/24)."DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs." Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression 781(1-2): 130-142. <http://hdl.handle.net/2027.42/24900>
http://www.sciencedirect.com/science/article/B6T1V-47S5YFF-H9/2/1028d1c25c4c921d5dc98e29cb8dbbe1
http://www.ncbi.nlm.nih.gov/sites/entrez?cmd=retrieve&db=pubmed&list_uids=6320890&dopt=citation
http://hdl.handle.net/2027.42/24900
6320890
http://dx.doi.org/10.1016/0167-4781(84)90131-3
Biochimica et Biophysica Acta
Appears in Collections:Research Collections

Files in This Item:
Click on the URI links for accessing contents.