Model for T-antigen-dependent melting of the simian virus 40 core origin based on studies of the interaction of the beta-hairpin with DNA.

Abstract:

:The interaction of simian virus 40 (SV40) T antigen (T-ag) with the viral origin has served as a model for studies of site-specific recognition of a eukaryotic replication origin and the mechanism of DNA unwinding. These studies have revealed that a motif termed the "beta-hairpin" is necessary for assembly of T-ag on the SV40 origin. Herein it is demonstrated that residues at the tip of the "beta-hairpin" are needed to melt the origin-flanking regions and that the T-ag helicase domain selectively assembles around one of the newly generated single strands in a manner that accounts for its 3'-to-5' helicase activity. Furthermore, T-ags mutated at the tip of the "beta-hairpin" are defective for oligomerization on duplex DNA; however, they can assemble on hybrid duplex DNA or single-stranded DNA (ssDNA) substrates provided the strand containing the 3' extension is present. Collectively, these experiments indicate that residues at the tip of the beta-hairpin generate ssDNA in the core origin and that the ssDNA is essential for subsequent oligomerization events.

journal_name

J Virol

journal_title

Journal of virology

authors

Kumar A,Meinke G,Reese DK,Moine S,Phelan PJ,Fradet-Turcotte A,Archambault J,Bohm A,Bullock PA

doi

10.1128/JVI.02451-06

subject

Has Abstract

pub_date

2007-05-01 00:00:00

pages

4808-18

issue

9

eissn

0022-538X

issn

1098-5514

pii

JVI.02451-06

journal_volume

81

pub_type

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