Abstract:
:The DNA unwinding element (DUE) is a sequence rich in adenine and thymine residues present within the origin region of both prokaryotic and eukaryotic replicons. Recently, it has been shown that this is the site where bacterial DnaA proteins, the chromosomal replication initiators, form a specific nucleoprotein filament. DnaA proteins contain a DNA binding domain (DBD) and belong to the family of origin binding proteins (OBPs). To date there has been no data on whether OBPs structurally different from DnaA can form nucleoprotein complexes within the DUE. In this work we demonstrate that plasmid Rep proteins, composed of two Winged Helix domains, distinct from the DBD, specifically bind to one of the strands of ssDNA within the DUE. We observed nucleoprotein complexes formed by these Rep proteins, involving both dsDNA containing the Rep-binding sites (iterons) and the strand-specific ssDNA of the DUE. Formation of these complexes required the presence of all repeated sequence elements located within the DUE. Any changes in these repeated sequences resulted in the disturbance in Rep-ssDNA DUE complex formation and the lack of origin replication activity in vivo or in vitro.
journal_name
Nucleic Acids Resjournal_title
Nucleic acids researchauthors
Wegrzyn K,Fuentes-Perez ME,Bury K,Rajewska M,Moreno-Herrero F,Konieczny Idoi
10.1093/nar/gku453subject
Has Abstractpub_date
2014-07-01 00:00:00pages
7807-18issue
12eissn
0305-1048issn
1362-4962pii
nar/gku453journal_volume
42pub_type
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