Abstract
Adeno-associated virus 2 (AAV), a nonpathogenic human virus, can either integrate into host chromatin and remain latent or replicate following infection. The outcome depends on the cellular conditions. Under conditions permissive for AAV DNA replication (e.g., during adenovirus coinfection), AAV gene expression is induced. The single-stranded AAV genome of 4680 nucleotides is organized into two open reading frames (ORFs) that encode structural capsid proteins (Cap) and nonstructural proteins (Rep) (Fig. 1). Two promoters at AAV map units 5 and 19, p5 and p19, direct expression of the rep gene. The cap gene is regulated by the p40 promoter. A common intron results in the production of four Rep proteins: p5 initiated Rep78, Rep68 and p19 initiated Rep52, Rep40. The inverted terminal repeats (ITRs) function as viral origins of replication required for encapsidation of the of AAV DNA. The production of AAV Rep proteins enables viral DNA to replicate, resulting in a geometric increase in the number of viral genomes. The AAV p5 initiated regulatory proteins Rep78 and Rep68 interact with the viral promoters to establish a feedback loop. These Rep proteins are involved directly in viral DNA replication as well.
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References
Ashktorab H, Srivastava A (1989) Identification of nuclear proteins that specifically interact with adeno-associated virus type 2 inverted terminal repeat hairpin DNA. J Virol 63: 3034–3039
Batchu RB, Kotin RM, Hermonat PL (1994) The regulatory rep protein of adeno-associated virus binds to sequences within the c-H-ras promoter. Cancer Lett 86: 23–31
Beaton A, Palumbo P, Berns KI (1989) Expression from the adeno-associated virus p5 and p19 promoters is negatively regulated in trans by the rep protein. J Virol 63: 4450–4454
Berns KI, Kotin RM, Labow MA (1988) Regulation of adeno-associated virus DNA replication. Biochim Biophys Acta 951: 425–429
Cheung AK, Hoggan MD, Hauswirth WW, Berns KI (1980) Integration of the adeno-associated virus genome into cellular DNA in latently infected human Detroit 6 cells. J Virol 33: 739–748
Chiorini JA, Weitzman MD, Owens RA, Urcelay E, Safer B, Kotin RM (1994a) Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli. J Virol 68: 797–804
Chiorini JA, Wiener SM, Owens RA, Kyostio SRM, Kotin RM, Safer B (1994b) Sequence requirements for stable binding and function of adeno-associated virus 2 Rep68 protein on the itr. J Virol 68: 7448–7457
Hermonat PL (1989) The adeno-associated virus Rep78 gene inhibits cellular transformation induced by bovine papillomavirus. Virology 172: 253–261
Hermonat PL (1991) Inhibition of H-ras expression by the adeno-associated virus Rep78 transformation suppressor gene product. Cancer Res 51: 3373–3377
Hermonat PL (1992) Inhibition of bovine papillomavirus plasmid DNA replication by adeno-associated virus. Virology 198: 329–333
Hermonat PL, Labow MA, Wright R, Berns KI, Muzyczka N (1984) Genetics of adeno-associated virus: isolation and preliminary characterization of adeno-associated virus type 2 mutants. J Virol 51: 329–339
Hong G, Ward P, Berns KI (1992) In vitro replication of adeno-associated virus DNA. Proc Natl Acad Sci USA 89: 4673–4677
Im DS, Muzyczka N (1989) Factors that bind to adeno-associated virus terminal repeats. J Virol 63: 3095–3104
Im D-S, Muzyczka N (1990) The AAV origin binding protein is an ATP-dependent site-specific endonuclease with DNA helicase activity. Cell 61: 447–457
Im DS, Muzyczka N (1992) Partial purification of adeno-associated virus Rep78, Rep52, and Rep40 and their biochemical characterization. J Virol 66: 1119–1128
Katze MG, Dorato D, Krug RM (1986) Cellular NA translation is blocked at both initiation and elongation after infection by influenza virus or adenovirus. J Virol 60: 1027–1039
Khleif SN, Myers T, Carter BJ, Trempe JP (1991) Inhibition of cellular transformation by the adeno-associated virus rep gene. Virology 181: 738–741
Kotin RM (1994) Prospects for the use of adeno-associated virus as a vector for human gene therapy. Hum Gene Ther 5: 793–801
Kotin RM, Siniscalco M, Samulski RJ, Zhu X, Hunter L, Laughlin CA, McLaughlin S, Muzyczka N, Rocchi M, Berns KI (1990) Site-specific integration by adeno-associated virus. Proc Nat Acad Sci USA 87: 2211–2215
Kotin RM, Menninger JC, Ward DC, Berns KI (1991) Mapping and direct visualization of a region-specific viral DNA integration site on chromosome 19q13-qter. Genomics 10: 831–834
Kotin RM, Linden RM, Berns KI (1992) Characterization of a preferred site on human chromosome 19q for integration of adeno-associated virus DNA by non-homologous recombination. EMBO J 11: 5071–5078
Krausslich HG, Nickün MJ, Toyoda H, Ethcison D, Wimmer E (1987) Poliovirus proteinase 2A induces cleavage of eukaryotic initiation factor 4F polypeptide p220. J Virol 61: 2711–2718
Kyostio S, Owens R, Weitzman M, Antoni B, Chejanovsky N, Carter B (1994) Analysis of adeno-associated (AAV) wild-type and mutant Rep proteins for their ability to regulate negatively AAV p5 and p19 NA levels. J Virol 68: 2947–2957
Labow MA, Berns KI (1988) The adeno-associated virus rep gene inhibits replication of an adeno-associated virus/simian virus 40 hybrid genome in cos-7 cells. Mol Cell Biol 7: 1320–1325
Labow MA, Hermonat PL, Berns KI (1986) Positive and negative autoregulation of the adeno-associated virus type 2 genome. J Virol 60: 515–524
Labow MA, Graf LH, Berns KI (1987) Adeno-associated virus gene expression inhibits cellular transformation by heterologous genes. Mol Cell Biol 7: 1320–1325
McCarty DM, Pereira DJ, Zolotukhin I, Zhou X, Ryan JH, Muzyczka N (1994a) Identification of linear DNA sequences that specifically bind the adeno-associated virus Rep protein. J Virol 68: 4988–4997
McCarty DM, Ryan JH, Zolotukhin S, Zhou X, Muzyczka N (1994b) Interaction of the adeno-associated virus Rep protein with a sequence within the A palindrome of the viral terminal repeat. J Virol 68: 4998–5006
McLaughlin SK, Collis P, Hermonat PL, Muzyczka N (1988) Adeno-associated virus general transduction vectors: analysis of proviral structures. J Virol 62: 1963–1973
Ni T-H, Zhou X, Marty D, Zolotukhin I, Muzyczka N (1994) In vitro replication of adeno-associated virus DNA. J Virol 68: 1128–1138
Oelze I, Rittner K, Sczakiel G (1994) Adeno-associated virus type 2 rep gene-mediated inhibition of basal expression of human immunodeficiency virus type 1 involves its negative regulatory functions. J Virol 68: 1229–1233
Owens RA, Carter BJ (1992) In vitro resolution of adeno-associated virus DNA hairpin termini by wild-type Rep protein is inhibited by a dominant-negative mutant of rep. J Virol 66: 1236–1240
Owens RA, Trempe JP, Chejanovsky N, Carter BJ (1991) Adeno-associated virus rep proteins produced in insect and mammalian expression systems: wild-type and dominant-negative mutant proteins bind to the viral replication origin. Virology 184: 14–22
Owens RA, Weitzman MD, Kyostio SR, Carter BJ (1993) Identification of a DNA-binding domain in the amino terminus of adeno-associated virus Rep proteins. J Virol 67: 997–1005
Pilder S, Moore M, Logan J, Shenk T (1986) The adenovirus E1B-55K transforming polypeptide modulates transport or cytoplasmic stabilization of viral and host cell NAs. Mol Cell Biol 6: 470–476
Samulski RJ, Zhu X, Xiao X, Brook JD, Housman DE, Epstein N, Hunter LA (1991) Targeted integration of adeno-associated virus (AAV) into human chromosome 19 (published erratum appears in EMBO J 1992 Mar; 11(3): 1228). EMBO J 10: 3941–3950
Shelling A, Smith M (1994) Targeted integration of transfected and infected adeno-associated virus vectors containing the neomycin resistance gene. Gene Ther 2: 1–5
Snyder R, Im D-S, Ni T, Xiao X, Samulski R, Muzyczka N (1993) Features of the adeno-associated virus origin involved in substrate recognition by the viral Rep protein. J Virol 67: 6096–6104
Srivastava A, Lusby EW, Berns KI (1983) Nucleotide sequence and organization of the adeno-associated virus 2 genome. J Virol 45: 555–564
Trempe JP, Carter BJ (1988) Regulation of adeno-associated virus gene expression in 293 cells: control of mRNA abundance and translation. J Virol 62: 68–74
Urcelay E, Ward P, Wiener SM, Safer B, Kotin RM (1995) Asymmetric replication in vitro from a human sequence element is dependent on adeno-associated virus Rep protein. J Virol 69 (in press)
Ward P, Berns KI (1991) In vitro rescue of an integrated hybrid adeno-associated virus/simian virus 40 genome. J Mol Biol 218: 791–804
Weitzman MD, Kyostio SRM, Kotin RM, Owens RA (1994) Adeno-associated virus (AAV) rep proteins mediate complex formation between AAV DNA and the human integration site. Proc Natl Acad Sci USA 91: 5808–5812
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© 1996 Springer-Verlag Berlin Heidelberg
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Chiorini, J.A. et al. (1996). The Roles of AAV Rep Proteins in Gene Expression and Targeted Integration. In: Berns, K.I., Giraud, C. (eds) Adeno-Associated Virus (AAV) Vectors in Gene Therapy. Current Topics in Microbiology and Immunology, vol 218. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-80207-2_2
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DOI: https://doi.org/10.1007/978-3-642-80207-2_2
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