Nucleic Acids Research, Vol 25, Issue 7 1347-1354, Copyright © 1997 by Oxford University Press
S Portes-Sentis, A Sergeant and H Gruffat
OriLyt, thecis-acting element of Epstein-Barr virus lytic origin of
replication, consists of upstream and downstream components. The upstream
component plays a dual role in transcription and replication. The
downstream component contains a homopurine-homopyrimidine sequence which
forms an H palindrome. We show that the downstream component can adopt a
triple helix structure in vitro, that the 5' border of the homopyrimidine
sequence is sensitive to P1 nuclease when carried by a supercoiled plasmid
and that an oligonucleotide complementary to the homopyrimidine strand is
taken up by a plasmid carrying the OriLyt H palindrome. We also show that
all mutations which alter the H palindrome impair both oligonucleotide
uptake and OriLyt-dependent replication. Interestingly, compensatory
mutations which restore an H palindrome also restore oligonucleotide uptake
by the mutated plasmids and their OriLyt-dependent replication. Thus, there
is a strong correlation between the inability of the OriLyt H palindrome to
form a non-B-DNA structure in vitro and impairment of OriLyt-dependent
replication. This suggests that the presence of a non-B-DNA structure in
the OriLyt downstream component is required for OriLyt-dependent
replication.
ARTICLES
A particular DNA structure is required for the function of a cis-acting component of the Epstein-Barr virus OriLyt origin of replication
Laboratoire de Virologie Humaine, U412 INSERM, ENS-Lyon, 46 Allee d'Italie, F-69364 Lyon cedex 07, France.
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