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Complementary intrastrand base pairing during initiation of Herpes simplex virus type 1 DNA replication.

Artikel i vetenskaplig tidskrift
Författare Alireza Aslani
Bertil Macao
Stina Simonsson
Per Elias
Publicerad i Proceedings of the National Academy of Sciences of the United States of America
Volym 98
Nummer/häfte 13
Sidor 7194-9
ISSN 0027-8424
Publiceringsår 2001
Publicerad vid Institutionen för medicinsk och fysiologisk kemi
Sidor 7194-9
Språk en
Länkar dx.doi.org/10.1073/pnas.121177198
Ämnesord Adenosine Triphosphate, metabolism, Animals, Base Pairing, Base Sequence, Cell Line, DNA Replication, DNA, Viral, chemistry, genetics, DNA-Binding Proteins, chemistry, genetics, metabolism, Herpesvirus 1, Human, genetics, Models, Molecular, Molecular Sequence Data, Nucleic Acid Conformation, Oligodeoxyribonucleotides, chemistry, Recombinant Proteins, chemistry, metabolism, Replication Origin, Spodoptera, Transfection, Viral Proteins, chemistry, genetics, metabolism
Ämneskategorier Virologi, Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)

Sammanfattning

The herpes simplex virus type 1 origin of DNA replication, oriS, contains three copies of the recognition sequence for the viral initiator protein, origin binding protein (OBP), arranged in two palindromes. The central box I forms a short palindrome with box III and a long palindrome with box II. Single-stranded oriS adopts a conformation, oriS*, that is tightly bound by OBP. Here we demonstrate that OBP binds to a box III-box I hairpin with a 3' single-stranded tail in oriS*. Mutations designed to destabilize the hairpin abolish the binding of OBP to oriS*. The same mutations also inhibit DNA replication. Second site complementary mutations restore binding of OBP to oriS* as well as the ability of mutated oriS to support DNA replication. OriS* is also an efficient activator of the hydrolysis of ATP by OBP. Sequence analyses show that a box III-box I palindrome is an evolutionarily conserved feature of origins of DNA replication from human, equine, bovine, and gallid alpha herpes viruses. We propose that oriS facilitates initiation of DNA synthesis in two steps and that OBP exhibits exquisite specificity for the different conformations oriS adopts at these stages. Our model suggests that distance-dependent cooperative binding of OBP to boxes I and II in duplex DNA is succeeded by specific recognition of a box III-box I hairpin in partially unwound DNA.

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