Identification of linear DNA sequences that specifically bind the adeno-associated virus Rep protein.

摘要: We have used baculovirus-expressed Rep68 that has been purified to homogeneity reexamine the binding properties of Rep protein. find is capable a linear DNA sequence contained within 25-bp A stem adeno-associated virus (AAV) terminal repeat proximal B and C palindromes. This shown conclusively by demonstrating could specifically bind synthetic oligonucleotide containing region in absence other sequences repeat. Rep78 was also recognition element, as demonstrated fact affinity column can be purify Rep78. The ability recognize provides mechanism which protein oriented on so only correct strand cut at resolution site (trs site) during resolution. In addition, computer analysis suggests similar element are present three AAV promoter regions. Electrophoretic mobility shift experiments clearly demonstrate p5 contains sequence. DNase protection indicate located between YY1 initiator TATA site. position immediately act repressor or transactivator transcription interacting with either TBP. gel suggest p19 presence sites upstream both promoters these may involved coordinate regulation transcription. we identified heterologous pBR322 DNA. comparison stem, p5, repeating GAGC motif least part accompanying report (D. M. McCarty, J. H. Ryan, S. Zolutukhin, X. Zhou, N. Muzyczka, Virol. 68:4998-5006, 1994), examine relative complete Finally, reexamined trs substrates do not contain palindromes any apparent secondary structure.(ABSTRACT TRUNCATED AT 400 WORDS)