Adenovirus DNA replication in vitro
- PMID: 284391
- PMCID: PMC383008
- DOI: 10.1073/pnas.76.2.655
Adenovirus DNA replication in vitro
Abstract
A soluble extract from the nuclei of HeLa cells infected with adenovirus 5 (Ad5) carries out the semiconservative replication of exogenously added Ad5 DNA in vitro. Maximal DNA synthesis is observed when DNA-protein complex, isolated from Ad5 virions, is added as template. DNA-protein complex from virions of the closely related virus, adenovirus 2, is also active. In contrast, very little in vitro DNA synthesis is observed when deproteinized Ad5 DNA or DNA from a heterologous source (bacteriophage T7) is added as template. The product of the in vitro reaction consists of long Ad5 DNA strands that are hydrogen-bonded, but not covalently linked, to the input DNA template. During the course of the in vitro reaction, branched molecules with structural features identical to in vivo replication intermediates are formed. These findings support the conclusion that replication in the in vitro system closely resembles adenovirus DNA replication in vivo. The system provides an assay that should be useful for the purification and subsequent characterization of viral and cellular proteins involved in DNA replication.
Similar articles
-
[Synthesis of adenovirus SA7 and Ad5 DNA in extracts from the nuclei of green monkey kidney cells].Mol Biol (Mosk). 1982 Jul-Aug;16(4):782-9. Mol Biol (Mosk). 1982. PMID: 7121461 Russian.
-
Complementation of the temperature-sensitive defect in H5ts125 adenovirus DNA replication in vitro.Proc Natl Acad Sci U S A. 1979 Nov;76(11):5534-8. doi: 10.1073/pnas.76.11.5534. Proc Natl Acad Sci U S A. 1979. PMID: 230480 Free PMC article.
-
Simian virus 40 DNA replication in vitro.Proc Natl Acad Sci U S A. 1984 Nov;81(22):6973-7. doi: 10.1073/pnas.81.22.6973. Proc Natl Acad Sci U S A. 1984. PMID: 6095264 Free PMC article.
-
Replication of origin containing adenovirus DNA fragments that do not carry the terminal protein.Nucleic Acids Res. 1983 Apr 11;11(7):1975-89. doi: 10.1093/nar/11.7.1975. Nucleic Acids Res. 1983. PMID: 6300787 Free PMC article.
-
Adenovirus DNA replication in vitro: a protein linked to the 5' end of nascent DNA strands.J Virol. 1981 Jan;37(1):139-47. doi: 10.1128/JVI.37.1.139-147.1981. J Virol. 1981. PMID: 7218423 Free PMC article.
Cited by
-
Adenovirus DNA replication in vitro: site-directed mutagenesis of the nuclear factor I binding site of the Ad2 origin.Nucleic Acids Res. 1985 Jul 11;13(13):4935-52. doi: 10.1093/nar/13.13.4935. Nucleic Acids Res. 1985. PMID: 4040630 Free PMC article.
-
Transcription initiation by RNA polymerase II does not require hydrolysis of the beta-gamma phosphoanhydride bond of ATP.EMBO J. 1994 Jan 15;13(2):391-9. doi: 10.1002/j.1460-2075.1994.tb06273.x. EMBO J. 1994. PMID: 8313884 Free PMC article.
-
Morphogenesis of bacteriophage phi 29 of Bacillus subtilis: oriented and quantized in vitro packaging of DNA protein gp3.J Virol. 1983 Jan;45(1):383-96. doi: 10.1128/JVI.45.1.383-396.1983. J Virol. 1983. PMID: 6185695 Free PMC article.
-
Adenovirus DNA replication in vitro: synthesis of full-length DNA with purified proteins.Proc Natl Acad Sci U S A. 1983 Jul;80(14):4266-70. doi: 10.1073/pnas.80.14.4266. Proc Natl Acad Sci U S A. 1983. PMID: 6308611 Free PMC article.
-
Processing of the adenovirus terminal protein.J Virol. 1981 Apr;38(1):272-7. doi: 10.1128/JVI.38.1.272-277.1981. J Virol. 1981. PMID: 7241655 Free PMC article.
References
Publication types
MeSH terms
LinkOut - more resources
Full Text Sources
Other Literature Sources
