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Microbiology 144 (1998), 1281-1290; DOI  10.1099/00221287-144-5-1281
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Structural elements of the Streptomyces oriC region and their interactions with the DnaA protein

Dagmara Jakimowicz1, Jerzy Majka1,2, Walter Messer2, Christian Speck2, Marisol Fernandez3, M Cruz Martin3, Jesus Sanchez3, Florian Schauwecker4, Ullrich Keller4, Hildgund Schrempf5 and Jolanta Zakrzewska-CzerwinAska1,*

Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wroclaw, Poland
Max-Planck-Institut für Molekulare Genetik, Ihnestraβe 73, D-14195 Berlin-Dahlem, Germany
Departamento de Biologia Funcional e Instituto Universitario de Biotecnologia de Asturias, Universidad de Oviedo, J. Claveria 6, Oviedo 33006, Spain
Max-Volmer-Institut für Biophysikalische Chemie und Biochemie, Fachgebiet Biochemie und Molekulare Biologie, Technische Universität Berlin, Franklinstrasse 29, D-10587 Berlin, Germany
Fachbereich Biologie/Chemie, Universität Osnabrück, Barbarastraβe 11, 49069 Osnabrück, Germany

ABSTRACT

Streptomycetes differ from other prokaryotic organisms in their mycelial life cycle and in possessing a large, linear, GC-rich chromosome. To deduce structural features of the Streptomyces origin of chromosomal replication, the oriC sequences of three Streptomyces species (S. antibioticus, S. chrysomallus and S. lividans) were compared. In Streptomyces, the oriC region contains 19 DnaA boxes whose location, orientation and spacing are conserved. The consensus sequence of the DnaA box identified within Streptomyces oriC is (T/C)(T/C)(G/A/C)TCCACA (preferred bases underlined). The interactions of DnaA with DNA fragments containing single, two or three DnaA boxes were studied using surface plasmon resonance. The dissociation constant (KD) for specific binding of individual DnaA boxes varied between 12 and 78 nM. Streptomyces oriC does not contain the three AT-rich 13-mer direct repeats present in the 5' part of the Escherichia coli oriC region. However, short AT-rich sequences are distributed among the DnaA boxes of Streptomyces oriC. Repeated attempts to unwind Streptomyces oriC have been unsuccessful. It remains to be elucidated whether DnaA interacts with putative accessory proteins which help in unwinding Streptomyces oriC.

*Author for correspondence: Jolanta Zakrzewska-CzerwinAska. Tel: +48 71 67 90 81. Fax: +48 71 67 91 11. e-mail: zakrzew@immuno.iitd.pan.wroc.pl

Keywords: DnaA box, AT-rich sequence, oriC, Streptomyces




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