Structural elements of the Streptomyces oriC region and their interactions with the DnaA protein.
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.
Jakimowicz
Dagmara
D
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wroclaw, Poland.
Majka
Jerzy
J
Max-Planck-Institut für Molekulare Genetik, Ihnestraße 73, D-14195 Berlin-Dahlem, Germany.|Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wroclaw, Poland.
Messer
Walter
W
Max-Planck-Institut für Molekulare Genetik, Ihnestraße 73, D-14195 Berlin-Dahlem, Germany.
Speck
Christian
C
Max-Planck-Institut für Molekulare Genetik, Ihnestraße 73, D-14195 Berlin-Dahlem, Germany.
Fernandez
Marisol
M
Departamento de Biologia Funcional e Instituto Universitario de Biotecnologia de Asturias, Universidad de Oviedo, J. Claveria 6, Oviedo 33006, Spain.
Cruz Martin
M
M
Departamento de Biologia Funcional e Instituto Universitario de Biotecnologia de Asturias, Universidad de Oviedo, J. Claveria 6, Oviedo 33006, Spain.
Sanchez
Jesus
J
Departamento de Biologia Funcional e Instituto Universitario de Biotecnologia de Asturias, Universidad de Oviedo, J. Claveria 6, Oviedo 33006, Spain.
Schauwecker
Florian
F
Max-Volmer-Institut für Biophysikalische Chemie und Biochemie, Fachgebiet Biochemie und Molekulare Biologie, Technische Universität Berlin, Franklinstrasse 29, D-10587 Berlin, Germany.
Keller
Ullrich
U
Max-Volmer-Institut für Biophysikalische Chemie und Biochemie, Fachgebiet Biochemie und Molekulare Biologie, Technische Universität Berlin, Franklinstrasse 29, D-10587 Berlin, Germany.
Schrempf
Hildgund
H
Fachbereich Biologie/Chemie, Universität Osnabrück, Barbarastraße 11, 49069 Osnabrück, Germany.
Zakrzewska-Czerwinńska
Jolanta
J
Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, ul. Weigla 12, 53-114 Wroclaw, Poland.
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004261:DNA Replication; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008969:Molecular Sequence Data; D012045:Regulatory Sequences, Nucleic Acid; D018741:Replication Origin; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D013302:Streptomyces / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C066312:DnaA protein, Bacteria