Architecture of the Streptomyces lividans DnaA protein-replication origin complexes

Autor(en): Jakimowicz, D
Majka, J
Konopa, G
Wegrzyn, G
Messer, W
Schrempf, H 
Zakrzewska-Czerwinska, J
Stichwörter: BARR NUCLEAR ANTIGEN-1; bending; Biochemistry & Molecular Biology; COELICOLOR CHROMOSOME; DIMER; dimerization; DnaA protein; ESCHERICHIA-COLI; GENE; INITIATION; LAMBDA; oriC; PLASMID; REGION; Streptomyces lividans; TARGET
Erscheinungsdatum: 2000
Volumen: 298
Ausgabe: 3
Startseite: 351
Seitenende: 364
The Streptomyces oriC region contains two clusters of 19 DnaA boxes separated by a spacer (134 bp). The Streptomyces DnaA protein consists, like all other DnaA proteins, of four domains: domain III and the carboxyterminal part (domain IV) are responsible for binding of ATP and DNA, respectively. Binding of the DnaA protein to the entire oriC region analysed by electron microscopy showed that the DnaA. protein forms separate complexes at each of the clusters of DnaA boxes, but not at the spacer separating them. In vivo mutational analysis revealed that the number of DnaA boxes and the presence of the spacer linking both groups of DnaA boxes seem to be important for a functional Streptomyces origin. We suggest that the arrangement of DnaA boxes allows the DNA-bound DnaA protein to induce bending and looping of the oriC region. As it was shown by electrophoretic mobility shift assay and ``one hybrid system'', two domains, I and III, facilitate interactions between DnaA molecules. We postulate that domain I and domain III could be involved in cooperativity at distant and at closely spaced DnaA boxes, respectively. The long domain II extends the range over which N termini (domain I) of DNA-bound DnaA protein can form dimers. Thus, interactions between DnaA molecules may bring two clusters of DnaA boxes separated by the spacer into functional contact by loop formation. Removal of the spacer region or deletion of domains I and II resulted, respectively, in nucleoprotein complexes which are not fully developed, or huge nucleoprotein aggregates. (C) 2000 Academic Press.
ISSN: 00222836
DOI: 10.1006/jmbi.2000.3686

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