DNA bending, compaction and negative supercoiling by the architectural protein Sso7d of Sulfolobus solfataricus.
Members of the Sso7d/Sac7d family are small, abundant, non-specific DNA-binding proteins of the hyperthermophilic Archaea SULFOLOBUS: Crystal structures of these proteins in complex with oligonucleotides showed that they induce changes in the helical twist and marked DNA bending. On this basis they have been suggested to play a role in organising chromatin structures in these prokaryotes, which lack histones. We report functional in vitro assays to investigate the effects of the observed Sso7d-induced structural modifications on DNA geometry and topology. We show that binding of multiple Sso7d molecules to short DNA fragments induces significant curvature and reduces the stiffness of the complex. Sso7d induces negative supercoiling of DNA molecules of any topology (relaxed, positively or negatively supercoiled) and in physiological conditions of temperature and template topology. Binding of Sso7d induces compaction of positively supercoiled and relaxed DNA molecules, but not of negatively supercoiled ones. Finally, Sso7d inhibits the positive supercoiling activity of the thermophile-specific enzyme reverse gyrase. The proposed biological relevance of these observations is that these proteins might model the behaviour of DNA in constrained chromatin environments.
Napoli
Alessandra
A
Institute of Protein Biochemistry and Enzymology, Consiglio Nazionale delle Ricerche, Via Pietro Castellino 111, 80131 Naples, Italy.
Zivanovic
Yvan
Y
Bocs
Chantal
C
Buhler
Cyril
C
Rossi
Mose'
M
Forterre
Patrick
P
Ciaramella
Maria
M
D019843:Archaeal Proteins; D026942:DNA Topoisomerases / Q000378:metabolism; D004264:DNA Topoisomerases, Type I; D019641:DNA, Archaeal / Q000737:chemistry* / Q000378:metabolism; D004270:DNA, Circular / Q000737:chemistry; D004278:DNA, Superhelical / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000378:metabolism* / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D009690:Nucleic Acid Conformation; D016944:Sulfolobus; D059004:Topoisomerase I Inhibitors
D019843:Archaeal Proteins; D019641:DNA, Archaeal; D004270:DNA, Circular; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; C093228:Sso7d protein, Sulfolobus; D059004:Topoisomerase I Inhibitors; D026942:DNA Topoisomerases; C041813:DNA reverse gyrase; D004264:DNA Topoisomerases, Type I