Functional analysis of cis- and trans-acting replication factors of porcine circovirus type 1.

Abstract:

:The replication proteins Rep and Rep' of porcine circovirus type 1 (PCV1) are both capable of introducing and resealing strand discontinuities at the viral origin of DNA replication in vitro underlying genome amplification by rolling-circle replication. The PCV1 origin of replication encompasses the minimal binding site (MBS) of the Rep and Rep' proteins and an inverted repeat with the potential to form a stem-loop. In this study, both elements of the PCV1 origin were demonstrated to be essential for viral replication in transfected cells. Furthermore, investigation of conserved amino acid motifs within Rep and Rep' proteins revealed that the mutation of motifs I, II, and III and of the GKS box interfered with viral replication. In vitro studies demonstrated that motifs I to III were essential for origin cleavage, while the GKS box was dispensable for the initiation of viral replication. A covalent link between Rep/Rep' and the DNA after origin cleavage was demonstrated, providing a mechanism for energy conservation for the termination of replication.

journal_name

J Virol

journal_title

Journal of virology

authors

Steinfeldt T,Finsterbusch T,Mankertz A

doi

10.1128/JVI.02420-06

subject

Has Abstract

pub_date

2007-06-01 00:00:00

pages

5696-704

issue

11

eissn

0022-538X

issn

1098-5514

pii

JVI.02420-06

journal_volume

81

pub_type

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