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Initiation of DNA replication in a eukaryotic rolling-circle replicon: identification of multiple DNA-protein complexes at the geminivirus origin.

Abstract：

:The mechanism of initiation of DNA replication in eukaryotic rolling- circle replicons is still poorly understood in molecular terms. Geminiviruses, a family of plant DNA viruses, which use this strategy during part of their replicative cycle, replicate in the nucleus and are amenable to molecular studies. Except for the virally encoded initiator protein (Rep), geminivirus DNA replication relies on cellular factors, likely interfering with cell cycle regulation of the infected cell. Here, we report the identification of three distinct DNA-protein complexes of the DNA replication initiator protein encoded by wheat dwarf geminivirus (WDV) within viral regulatory sequences controlling DNA replication and transcription. We have mapped the WDV Rep binding sites by combining gel-shift assays, electron microscopy and DNase I footprinting. Two of the Rep-DNA complexes (C and the V) are high-affinity complexes, located in the proximity of the two divergent TATA boxes, at 150 and 90 bp, respectively, from the DNA replication initiation site. The third one, the O-complex, is a low-affinity complex, which can assemble under conditions supporting the DNA cleavage reaction. This suggests that it might be responsible for initiation of rolling-circle DNA replication in WDV and other members of the Mastrevirus genus.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Castellano MM,Sanz-Burgos AP,Gutiérrez C

doi

10.1006/jmbi.1999.2916

keywords:

subject

Has Abstract

pub_date

1999-07-16 00:00:00

pages

639-52

issue

3

eissn

0022-2836

issn

1089-8638

pii

S0022-2836(99)92916-9

journal_volume

290

pub_type

杂志文章

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