SUMO-1 modification regulates the protein stability of the large regulatory protein Rep78 of adeno associated virus type 2 (AAV-2)

The large Rep proteins Rep78 and Rep68 of the helper-dependent adeno associated virus type 2 (AAV-2) are essential for both site-specific integration of AAV DNA in the absence of helpervirus and productive AAV replication in the presence of helpervirus. We have identified UBC9, the E2 conjugating en...

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Veröffentlicht in:Virology (New York, N.Y.) N.Y.), 2004-12, Vol.330 (1), p.284-294
Hauptverfasser: Weger, Stefan, Hammer, Eva, Heilbronn, Regine
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container_title Virology (New York, N.Y.)
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creator Weger, Stefan
Hammer, Eva
Heilbronn, Regine
description The large Rep proteins Rep78 and Rep68 of the helper-dependent adeno associated virus type 2 (AAV-2) are essential for both site-specific integration of AAV DNA in the absence of helpervirus and productive AAV replication in the presence of helpervirus. We have identified UBC9, the E2 conjugating enzyme for the small ubiquitin-related polypeptide SUMO-1, as binding partner of the large Rep proteins in yeast two-hybrid analysis and in GST pulldown assays. Modification of the large Rep proteins with SUMO-1 could be demonstrated in immunoblot analysis and in immunoprecipitations, with the lysine residue at amino acid position 84 serving as the major attachment site. The largely sumolation-deficient Rep78 lysine to arginine point mutant showed a strongly reduced half-life as compared to the wild-type protein. This finding implicates a role for sumolation in the regulation of Rep78 protein stability that is assumed to be critical for the establishment and maintenance of AAV latency.
doi_str_mv 10.1016/j.virol.2004.09.028
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source MEDLINE; Elsevier ScienceDirect Journals Complete; EZB-FREE-00999 freely available EZB journals
subjects AAV
Adeno associated
Adeno-associated virus 2
Cloning, Molecular
Dependovirus - metabolism
DNA Helicases - metabolism
DNA, Viral - genetics
DNA, Viral - isolation & purification
DNA-Binding Proteins - genetics
DNA-Binding Proteins - metabolism
Glutathione Transferase - metabolism
HeLa Cells
Humans
Kinetics
Latency
Modification
Open Reading Frames
Plasmids
Recombinant Fusion Proteins - metabolism
Rep
Saccharomyces cerevisiae - genetics
Saccharomyces cerevisiae - metabolism
Stability
SUMO
SUMO-1 Protein - metabolism
Viral Proteins - genetics
Viral Proteins - metabolism
title SUMO-1 modification regulates the protein stability of the large regulatory protein Rep78 of adeno associated virus type 2 (AAV-2)
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