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Abstract
The transcription system of archaea has both bacterial-like and eukaryotic-like features; however, only a small number of archaeal transcriptional regulators have been characterized in detail. This work describes the discovery and characterization of archaeal transcriptional regulators of the hyperthermophile Pyrococcus furiosus. A DNA affinity protein capture experiment was employed to pull specific DNA-binding proteins out of P. furiosus cell extract with promoter DNA. These proteins are responsible for the observed gene regulation in DNA microarray expression profiles of the transcriptional response of P. furiosus to cold shock. A novel transcription regulatory protein PF1072 was identified and its specific DNA binding ability at sites upstream of multiple genes was confirmed by the electrophoretic mobility shift assay and fluorescence-detected DNase I footprinting. Systematic evolution of ligands by exponential enrichment (SELEX) was used to determine the consensus binding sequence, so other binding sites in the genome can be detected and verified. The work presented here demonstrates that the novel transcriptional regulator PF1072 is likely a relevant participant in transcriptional regulation pathways related to P. furiosus cold shock adaptation.