Adenovirus DNA

The calcium phosphate method was first used in 1973 to introduce adenovirus DNA into mammalian cells [3]. DNA-Calcium-phosphate complexes are formed by mixing DNA in a phosphate buffer with calcium chloride. These complexes adhere to the cell membrane and enter the cytoplasm by endocytosis. Disadvantages of DEAE-dextran and calcium phosphate transfection are a certain level of cytotoxicity, a complicated transfection procedure, and the fact that not all cell types can be transfected using these methods. 

Trotman, L.C. et al. (2001) Import of adenovirus DNA involves the nuclear pole complex receptor CAN/ Nup214 and histone HI. Nat. Cell Biol. 3, 1092-1100. 

The spectrum of pleiotropic functions of different HI variants in different taxa has yet to be established. With respect to interactions within chromatin, the elucidation of the role of linker histones in transmitting or enhancing the effects of other non-histone proteins will be of particular interest. Given the dynamic behavior of HI in the nucleus, the possibility of auxiliary extrachromosomal functions performed by the free HI pool should be explored. An example of such a function is the recent finding that free HI is involved in the import of adenovirus DNA into the nucleus [158]. Other potential functions, especially at the period of maximum chromatin condensation, could depend on the gradient of free HI around the chromosomes. 

The role of the nuclear pore complex in adenovirus DNA entry. Embo J, 1997. 16(19) 5998-6007. 

Figure 28-13 (A) Stereoscopic ribbon drawing of the phyloge-netically conserved 180-residue C-terminal portion of the TATA-binding protein (TBP) from Arabidopsis thaliana. The sequence consists of two direct repeats, giving the protein an approximate twofold symmetry. From Nikolov et al.337 (B) Structure of the corresponding C-terminal core (residues 155-335) of the human TATA-binding protein (TBP) bound to the TATA sequence of a promoter in adenovirus DNA. From Nikolov et al.327 (C) Structure of human transcription factor IIB bound to a TBP from Arabidopsis thaliana, which, in turn, is bound to an adenovirus TATA sequence. Hypothetical B DNA extensions have been modeled at both ends of the DNA segment. The +1 at the left end is the transcription start site and the —43 upstream end is to the right. From Nikolov et al.338 Courtesy of Stephen K. Burley.

Chang, L. S. and Shenk, T. (1990). The adenovirus DNA-binding protein stimulates the rate of transcription directed by adenovirus and adeno-associated virus promoters. J. Virol. 64, 2103-2109. 

Janik, J. E., Huston, M. M., Cho, K. and Rose, J. A. (1989). Efficient synthesis of adeno-associated virus structural proteins requires both adenovirus DNA binding protein and VA I RNA. Virology 168, 320-329. 

Jay, F. T., Laughlin, C. A. and Carter, B. J. (1981). Eukaryotic translational control Adeno-associated virus protein synthesis is affected by a mutation in the adenovirus DNA-binding protein. Proc. Natl. Acad. Sci. USA 78, 2927-2931. 

Lithium (at 40 mM) inhibits the replication of herpes virus, pox virus, and adenovirus (DNA viruses) but does not inhibit that of RNA viruses such as influenza virus or encephalomyocarditis virus (209). 

The most extensively used adenoviruses are serotypes 2 (Ad2) and 5 (Ad5) because both are not associated with serious infectious disease in humans. - Similar to retroviral vectors, elements of adenovirus DNA genome are removed to prevent replication once inside the... 

Cristiano, R. J., Smith, L. C., Kay, M. A., Brinkely, B. R., and Woo, S. L. (1993) Hepatic gene therapy efficient gene delivery and expression in primary hepato-cytes utilizing a conjugated adenovirus-DNA complex. Proc. Natl. Acad. Sci. USA 90(24), 11,548-11,552. 

The precise base pair where RNA polymerase II initiates transcription in the adenovirus late transcription unit was determined by analyzing the RNAs synthesized during in vitro transcription of adenovirus DNA restriction fragments that extended somewhat upstream and downstream of the approximate initiation region determined by nascent-transcript analysis. The rationale of this experiment and typical results are illustrated in Figure 11-8. The RNA transcripts synthesized in vitro by RNA polymerase II from the... 

FIGURE 5.2 A single-strand circle formed by intra-molecular annealing of adenovirus DNA... 

FIGURE 5.3 Another proposal for formation of single-stranded molecules of adenovirus DNA. 

Horvath, 1., Palkonyay, L., and Weber, 1. 1986. Group C adenovirus DNA sequences in human lymphoid cells. J. Virol. 59, 189-192. 

Kolesar, J.M., Miller, J.A., Dhurandhar, N.V., and Atkinson, R.L. 2000. Direct quantification of AD-36 adenovirus DNA by capillary electrophoresis with laser-induced fiuorescence. J. Chromatogr. B Biomed Sci. Appl. 744, 1-8. 

Greber U, Suomalainen M, Stidwill R, Boucke K, Ebersold M, Helenius A (1997) The role of the nuclear pore complex in adenovirus DNA entry. EMBO J 16 5998-6007 Hale T, Braithwaite A (1999) The adenovirus oncoprotein Ela stimulates binding of transcription factor ETE to transcriptionally activate the p53 gene. J Biol Chem 274 23777-23786 Harty JT, Tvinnereim AR, White DW (2000) CD8 + T cell effector mechanisms in resistance to infection. Annu Rev Immunol 18 275-308... 

While the poly (A) sequences do seem to be involved in the transport of mRNA s from the nucleus, this does not seem to be the sole function of the poly (A) tract for example, adenovirus DNA appears to lack a DNA sequence complementary to poly (A) but replicates in the nucleus of the mammalian cell and appears to have a poly (A) tract added to the viral mRNA by host-cell mechanisms for transport of the adenovirus mRNA to the cytoplasm (Philipson et ah, 1971). As with cellular messages, cordycepin blocks both the labeling of the poly (A) tracts and the appearance of adenovirus-specific RNA in the cytoplasm of infected cells (Philipson et ah, 1971). In contrast, vaccinia virus replicates exclusively in the cytoplasm of cells it infects and still contains poly(A) sequences (Kates, 1970). Since no role in transport is involved here, it suggests that some mRNAs may require a poly (A) sequence for proper translation. Further, not all mammalian mRNAs contain poly (A) and still are transported to the cytoplasm for translation. Specifically, the 9 S histone message isolated by Adesnik and Darnell (1972) from HeLa cells lacks any detectable poly (A) sequence of any significant length. These workers have also shown that the exit time of the histone mRNA molecule from the nucleus is shorter than that of other messenger RNA s. 

Rosenfeld and Kelly (1) purified Nuclear Factor-I (NF-I), a Hela cell protein that enhances initiation of adenovirus DNA replication vitro, by affinity chromatography on a specific DNA-cellulose column. The column was prepared by adsorption to cellulose of a plasmid engineered to contain multiple (88) copies of the binding site for NF-1. NF-1 was purified from a Hela cell nuclear extract by chromatography on Bio-rex 70, coli DNA-cellulose, and two sequential passes on the specific DNA affinity column. 

Fig.1. Viruses. Schematic representation of virus particles, all drawn to scale. The type of genome (RNA or DNA) is shown in brackets. Enveloped viruses 1 Pox virus (DNA). 2 Rabies virus (RNA). 3 Influenza virus (RNA). 4 Measles virus (RNA). 5 Chickenpox virus (ONA). Naked or unenveloped viruses 6 Yellow fever virus (RNA). 7 Adenovirus (DNA). 8 Reovirus (RNA). 9 Wart-papilloma virus (ONA). 10 Poliomyelitis virus (RNA). 11 Parvovirus (RNA). 12 Corona virus (RNA). 13 Tobacco mosaic virus (RNA). 14 Bacteriophage T2 (DNA).

Yamada et al., 2002). Interestingly, Marin et al. (2000) showed that adenovirus DNA was present in the nasal passages of 78.4% of asthmatic children, but in only 5% of the healthy controls this suggested a link between adenovirus infection and asthma. 

Stunnenberg, H. G., Lange, H., Philipson, L., van Miltenburg, R. T., and van der Vliet, P. C. (1988) High expression of functional adenovirus DNA polymerase and precursor terminal protein using recombinant vaccinia virus. Nucleic Acids Res. 16, 2431-2444. 

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