Biochemical characterization of DNA-binding proteins from Pyrobaculum aerophilum and Aeropyrum pernix |
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Authors: | Christine D Hardy Patrick K Martin |
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Institution: | (1) Applied Biosystems, Inc., 850 Lincoln Centre Drive, MS 404-1, Foster City, CA 94404, USA |
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Abstract: | Several representatives of the Crenarchaeal branch of the Archaea contain highly abundant, small, positively charged proteins
exemplified by the Sso7d protein from Sulfolobus solfataricus. These proteins bind to DNA in a non-sequence-specific manner. Using publicly available genomic sequence information, we
identified a second class of small Crenarchaeal DNA-binding proteins represented by the Pyrobaculum aerophilum open reading frame 3192–encoded (Pae3192) protein and its paralogs. We investigated the biochemical properties of the Pae3192
protein and an orthologous protein (Ape1322b) from Aeropyrum pernix in side-by-side experiments with the Sso7d protein. We demonstrate that the recombinant Ape1322b, Pae3192 and Sso7d proteins
bind to DNA and that the DNA-protein complexes formed are slightly different for each protein. We show that like Sso7d, Pae3192
constrains negative supercoils in DNA. In addition, we show that all three proteins raise the melting temperature of duplex
DNA upon binding. Finally, we present the equilibrium affinity constants and kinetic association constants of each protein
for single-stranded and double-stranded DNA. |
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Keywords: | Archaea Crenarchaea Chromatin DNA-binding protein Thermostable Hyperthermophile |
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