scholarly journals Identification of nuclear proteins that specifically interact with adeno-associated virus type 2 inverted terminal repeat hairpin DNA.

1989 ◽  
Vol 63 (7) ◽  
pp. 3034-3039 ◽  
Author(s):  
H Ashktorab ◽  
A Srivastava
Keyword(s):  
Virus Type ◽  
Terminal Repeat ◽  
Nuclear Proteins ◽  
Inverted Terminal Repeat ◽  
Hairpin Dna ◽  
Adeno Associated Virus

scholarly journals Adeno-Associated Virus Type 2 p5 Promoter: a Rep-Regulated DNA Switch Element Functioning in Transcription, Replication, and Site-Specific Integration

2007 ◽  
Vol 81 (8) ◽  
pp. 3721-3730 ◽  
Author(s):  
Mary Murphy ◽  
Janette Gomos-Klein ◽  
Marko Stankic ◽  
Erik Falck-Pedersen
Keyword(s):  
Virus Type ◽  
Transcriptional Control ◽  
Helper Virus ◽  
Inverted Terminal Repeat ◽  
Sequence Element ◽  
Adeno Associated Virus ◽  
Site Specific ◽  
Site Specific Integration ◽  
Rep Proteins

ABSTRACT The large Rep proteins, p68 and p78, function as master controllers of the adeno-associated virus type 2 (AAV2) life cycle, involved in transcriptional control, in latency, in rescue, and in viral DNA replication. The p5 promoter may be the nucleic acid complement to the large Rep proteins. It drives expression of the large Rep proteins, it undergoes autoregulation by Rep, it undergoes induction by helper virus, it is a target substrate for Rep-mediated site-specific integration (RMSSI), and it can function as a replicative origin. To better understand the relationship between each of the p5 functions, we have determined the effects of p5 promoter mutations (p5 integration efficiency element, or p5IEE) on transcription, integration, and replication using RMSSI transfection protocols in HeLa cells. The data demonstrate that the organization of the p5 promoter provides a unique platform for regulated AAV2 template transcription and subsequent repression by Rep through direct and indirect mechanisms. The elements of the p5IEE that define its function as a promoter also define its function as a highly optimized substrate for Rep-mediated site-specific integration and replication. The p5 Rep binding element (RBE) is essential in RMSSI and Rep-dependent replication; however, replacement of the p5 RBE with either the AAV2 inverted terminal repeat or the AAVS1 RBE sequence elements neither enhances nor severely compromises RMSSI activity of p5IEE. The RBE by itself or in combination with the YY1+1 initiator/terminal resolution sequence element does not mediate efficient site-specific integration. We found that replication and integration were highly sensitive to sequence manipulations of the p5 TATA/RBE/YY1+1 core structure in a manner that reflects the function of these elements in transcription. The data presented support a model where, depending on the state of the cell (Rep expression and helper virus influences), the p5IEE operates as a transcription/integration switch sequence element.

scholarly journals Preferential Integration of Adeno-Associated Virus Type 2 into a Polypyrimidine/Polypurine-Rich Region within AAVS1

2007 ◽  
Vol 81 (18) ◽  
pp. 9718-9726 ◽  
Author(s):  
Victor J. McAlister ◽  
Roland A. Owens
Keyword(s):  
Virus Type ◽  
Virus Genome ◽  
Cellular Protein ◽  
Nonhomologous End Joining ◽  
Inverted Terminal Repeat ◽  
End Joining ◽  
Adeno Associated Virus ◽  
Group A ◽  
Xeroderma Pigmentosum Group A

ABSTRACT Adeno-associated virus type 2 (AAV2) preferentially integrates its genome into the AAVS1 locus on human chromosome 19. Preferential integration requires the AAV2 Rep68 or Rep78 protein (Rep68/78), a Rep68/78 binding site (RBS), and a nicking site within AAVS1 and may also require an RBS within the virus genome. To obtain further information that might help to elucidate the mechanism and preferred substrate configurations of preferential integration, we amplified junctions between AAV2 DNA and AAVS1 from AAV2-infected HeLaJW cells and cells with defective Artemis or xeroderma pigmentosum group A genes. We sequenced 61 distinct junctions. The integration junction sequences show the three classical types of nonhomologous-end-joining joints: microhomology at junctions (57%), insertion of sequences that are not normally contiguous with either the AAV2 or the AAVS1 sequences at the junction (31%), and direct joining (11%). These junctions were spread over 750 bases and were all downstream of the Rep68/78 nicking site within AAVS1. Two-thirds of the junctions map to 350 bases of AAVS1 that are rich in polypyrimidine tracts on the nicked strand. The majority of AAV2 breakpoints were within the inverted terminal repeat (ITR) sequences, which contain RBSs. We never detected a complete ITR at a junction. Residual ITRs at junctions never contained more than one RBS, suggesting that the hairpin form, rather than the linear ITR, is the more frequent integration substrate. Our data are consistent with a model in which a cellular protein other than Artemis cleaves AAV2 hairpins to produce free ends for integration.

Biologically active Rep proteins of adeno-associated virus type 2 produced as fusion proteins in Escherichia coli.

1994 ◽  
Vol 68 (2) ◽  
pp. 797-804 ◽  
Author(s):  
J A Chiorini ◽  
M D Weitzman ◽  
R A Owens ◽  
E Urcelay ◽  
B Safer ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virus Type ◽  
Fusion Proteins ◽  
Biologically Active ◽  
Adeno Associated Virus ◽  
Rep Proteins

scholarly journals Biodistribution of adeno‐associated virus type 2 carrying multi‐characteristic opsin in dogs following intravitreal injection

2021 ◽  
Vol 25 (18) ◽  
pp. 8676-8686
Author(s):  
Kissaou T. Tchedre ◽  
Subrata Batabyal ◽  
Melissa Galicia ◽  
Darryl Narcisse ◽  
Sourajit Mitra Mustafi ◽  
...  
Keyword(s):  
Intravitreal Injection ◽  
Virus Type ◽  
Adeno Associated Virus

scholarly journals Adeno-Associated Virus Type 2 and Hepatocellular Carcinoma?

2015 ◽  
Vol 26 (12) ◽  
pp. 779-781 ◽  
Author(s):  
Kenneth I. Berns ◽  
Barry J. Byrne ◽  
Terence R. Flotte ◽  
Guangping Gao ◽  
William W. Hauswirth ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Virus Type ◽  
Adeno Associated Virus

Induction of an Antitumoral Immune Response by Wild-Type Adeno-Associated Virus Type 2 in an In Vivo Model of Pancreatic Carcinoma

Pancreas ◽  
2007 ◽  
Vol 35 (1) ◽  
pp. 63-72 ◽  
Author(s):  
Sven Eisold ◽  
Jan Schmidt ◽  
Eduard Ryschich ◽  
Michael Gock ◽  
Ernst Klar ◽  
...  
Keyword(s):  
Immune Response ◽  
Pancreatic Carcinoma ◽  
Virus Type ◽  
In Vivo Model ◽  
Wild Type ◽  
Adeno Associated Virus

scholarly journals Presence of atrs-Like Motif Promotes Rep-Mediated Wild-Type Adeno-Associated Virus Type 2 Integration

2015 ◽  
Vol 89 (14) ◽  
pp. 7428-7432 ◽  
Author(s):  
Karl Petri ◽  
Richard Gabriel ◽  
Leticia Agundez ◽  
Raffaele Fronza ◽  
Saira Afzal ◽  
...  
Keyword(s):  
Virus Type ◽  
Integration Site ◽  
Dermal Fibroblasts ◽  
Wild Type ◽  
Mobility Shift ◽  
Adeno Associated Virus ◽  
Rep Protein ◽  
Electrophoretic Mobility Shift Assays ◽  
First Time

High-throughput integration site (IS) analysis of wild-type adeno-associated virus type 2 (wtAAV2) in human dermal fibroblasts (HDFs) and HeLa cells revealed that juxtaposition of a Rep binding site (RBS) and terminal resolution site (trs)-like motif leads to a 4-fold-increased probability of wtAAV integration. Electrophoretic mobility shift assays (EMSAs) confirmed binding of Rep to off-target RBSs. For the first time, we show Rep protein off-target nicking activity, highlighting the importance of the nicking substrate for Rep-mediated integration.

scholarly journals Charge-to-Alanine Mutagenesis of the Adeno-Associated Virus Type 2 Rep78/68 Proteins Yields Temperature-Sensitive and Magnesium-Dependent Variants

2000 ◽  
Vol 74 (1) ◽  
pp. 591-591
Author(s):  
Denise K. Gavin ◽  
Samuel M. Young ◽  
Weidong Xiao ◽  
Brenda Temple ◽  
Corinne R. Abernathy ◽  
...  
Keyword(s):  
Virus Type ◽  
Temperature Sensitive ◽  
Adeno Associated Virus
Sign in / Sign up

Export Citation Format

Share Document