Datensatz

Zusammenfassung

KorB is a member of the ParB family of bacterial partitioning proteins. The protein encoded by the conjugative plasmid RP4 is part of the global control circuit and regulates the expression of plasmid genes, the products of which are involved in replication, transfer, and stable inheritance. KorB is a homodimeric protein which binds to palindromic 13 bp DNA sequences [5'-TTTAGC(G/C)GCTAAA-3'] present 12 times in the 60 kb plasmid. Each KorB subunit is composed of two domains; the C-domain is responsible for the dimerization of the protein, whereas the N-terminal domain recognizes and binds to the operator sequence (OB). Here we describe results of a Raman spectroscopic study of the interaction of the N-domain with a double-stranded model oligonucleotide composed of the palindromic binding sequence and terminal 5'-ABrU and AG-3' bases. Comparison of the Raman spectra of the free KorB N-domain and OB DNA with the spectrum of the complex reveals large differences. KorB-N binds in the major groove of the OB DNA, and the interactions induce changes in the DNA backbone and in the secondary structure of the protein.