In vitro DNA binding of the archaeal protein Sso7d induces negative supercoiling at temperatures typical for thermophilic growth.
The topological state of DNA in hyperthermophilic archaea appears to correspond to a linking excess in comparison with DNA in mesophilic organisms. Since DNA binding proteins often contribute to the control of DNA topology by affecting DNA geometry in the presence of DNA topoisomerases, we tested whether the histone-like protein Sso7d from the hyperthermophilic archaeon Sulfolobus solfataricus alters DNA conformation. In ligase-mediated supercoiling assays carried out at 37, 60, 70, 80 and 90 degrees C we found that DNA binding of increasing amounts of Sso7d led to a progressive decrease in plasmid linking number (Lk), producing negative supercoiling. Identical unwinding effects were observed when recombinant non-methylated Sso7d was used. For a given Sso7d concentration the DNA unwinding induced was augmented with increasing temperature. However, after correction for the overwinding effect of high temperature on DNA, plasmids ligated at 60-90 degrees C exhibited similar sigma values at the highest Sso7d concentrations assayed. These results suggest that Sso7d may play a compensatory role in vivo by counteracting the overwinding effect of high temperature on DNA. Additionally, Sso7d unwinding could be involved in the topological changes observed during thermal stress (heat and cold shock), playing an analogous role in crenarchaeal cells to that proposed for HU in bacteria.
López-García
P
P
Institut de Génétique et Microbiologie, Université Paris-Sud, CNRS URA 1354, GDR 1006, Bâtiment 409, 91405 Orsay Cedex, France.
Knapp
S
S
Ladenstein
R
R
Forterre
P
P
D019843:Archaeal Proteins / Q000737:chemistry* / Q000378:metabolism; D019641:DNA, Archaeal / Q000737:chemistry* / Q000378:metabolism; D004278:DNA, Superhelical / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli; D006358:Hot Temperature; D008745:Methylation; D009690:Nucleic Acid Conformation; D010957:Plasmids / Q000737:chemistry; D011994:Recombinant Proteins; D016944:Sulfolobus / Q000235:genetics / Q000254:growth & development*
D019843:Archaeal Proteins; D019641:DNA, Archaeal; D004278:DNA, Superhelical; D004268:DNA-Binding Proteins; D011994:Recombinant Proteins; C093228:Sso7d protein, Sulfolobus