DNA-binding activity of adeno-associated virus Rep is required for inverted terminal repeat-dependent complex formation with herpes simplex virus ICP8.
Herpes simplex virus (HSV) helper functions for (AAV) replication comprise HSV ICP8 and helicase-primase UL5/UL52/UL8. Here we show that N-terminal amino acids of AAV Rep78 that contact the Rep-binding site within the AAV inverted terminal repeat (ITR) are required for ternary-complex formation with infected-cell protein 8 (ICP8) on AAV single-strand DNA (ssDNA) in vitro and for colocalization in nuclear replication domains in vivo. Our data suggest that HSV-dependent AAV replication is initiated by Rep contacting the AAV ITR and by cooperative binding of ICP8 on AAV ssDNA.
Institute of Virology, Campus Benjamin Franklin, Charité—Medical School, Berlin, Germany.
Martin
M
Alex
Stefan
S
Weger
Mario
M
Mietzsch
Heiko
H
Slanina
Toni
T
Cathomen
Regine
R
Heilbronn
D001665:Binding Sites; D004265:DNA Helicases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D000229:Dependovirus / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D006367:HeLa Cells; D006378:Helper Viruses / Q000235:genetics / Q000378:metabolism; D006801:Humans; D010322:Parvoviridae Infections / Q000821:virology; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D018139:Simplexvirus / Q000235:genetics / Q000378:metabolism*; D020079:Terminal Repeat Sequences; D014764:Viral Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D014779:Virus Replication
D004268:DNA-Binding Proteins; C045462:ICP8 protein, Simplexvirus; D014764:Viral Proteins; D004265:DNA Helicases