Crystal structure of the Lrp-like transcriptional regulator from the archaeon Pyrococcus furiosus

• Published in: The EMBO journal Vol. 20; no. 5; pp. 990 - 997
• Main Authors: Leonard, Philip M., Smits, Sander H.J., Sedelnikova, Svetlana E., Brinkman, Arie B., de Vos, Willem M., van der Oost, John, Rice, David W., Rafferty, John B.
• Format: Journal Article
• Language: English
• Published: Chichester, UK John Wiley & Sons, Ltd 01-03-2001; Blackwell Publishing Ltd; Oxford University Press
• Subjects: Amino Acid Sequence; Archaeal Proteins - chemistry; Archaeal Proteins - genetics; Archaeal Proteins - metabolism; AsnC protein; Binding Sites; Calorimetry, Differential Scanning; Crystallography, X-Ray; Deoxyribonucleic acid; Dimerization; DNA; DNA - chemistry; DNA - metabolism; DNA-Binding Proteins - chemistry; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; E coli; Escherichia coli Proteins; Gene Expression Regulation, Archaeal; helix-turn-helix; Laboratorium voor Microbiologie; Leucine - genetics; Leucine - metabolism; Leucine-Responsive Regulatory Protein; Lrp protein; Lrp-AsnC family; LrpA protein; Microbiological Laboratory; Microbiologie; Microbiology; Models, Molecular; Molecular Sequence Data; Mutation; Nucleic Acid Conformation; Protein Binding; Protein Structure, Secondary; Protein Structure, Tertiary; Pyrococcus furiosus; Pyrococcus furiosus - chemistry; Pyrococcus furiosus - genetics; Sequence Alignment; Transcription Factors - chemistry; Transcription Factors - genetics; Transcription Factors - metabolism; transcriptional regulator; VLAG; X-ray crystallography
• ISSN: 0261-4189; 1460-2075; 1460-2075
• DOI: 10.1093/emboj/20.5.990

The LrpA protein from the hyperthermophilic archaeon Pyrococcus furiosus belongs to the Lrp/AsnC family of transcriptional regulatory proteins, of which the Escherichia coli leucine‐responsive regulatory protein is the archetype. Its crystal structure has been determined at 2.9 Å resolution and is the first for a member of the Lrp/AsnC family, as well as one of the first for a transcriptional regulator from a hyperthermophile. The structure consists of an N‐terminal domain containing a helix–turn–helix (HtH) DNA‐binding motif, and a C‐terminal domain of mixed α/β character reminiscent of a number of RNA‐ and DNA‐binding domains. Pyrococcus furiosus LrpA forms a homodimer mainly through interactions between the antiparallel β‐sheets of the C‐terminal domain, and further interactions lead to octamer formation. The LrpA structure suggests how the protein might bind and possibly distort its DNA substrate through use of its HtH motifs and control gene expression. A possible location for an effector binding site is proposed by using sequence comparisons with other members of the family coupled to mutational analysis.