title: Structural basis for DNA-mediated allosteric regulation facilitated by the AAA+ module of Lon protease.
abstract: Lon belongs to a unique group of AAA+ proteases that bind DNA. However, the DNA-mediated regulation of Lon remains elusive. Here, the crystal structure of the α subdomain of the Lon protease from Brevibacillus thermoruber (Bt-Lon) is presented, together with biochemical data, and the DNA-binding mode is delineated, showing that Arg518, Arg557 and Arg566 play a crucial role in DNA binding. Electrostatic interactions contributed by arginine residues in the AAA+ module are suggested to be important to DNA binding and allosteric regulation of enzymatic activities. Intriguingly, Arg557, which directly binds DNA in the α subdomain, has a dual role in the negative regulation of ATPase stimulation by DNA and in the domain-domain communication in allosteric regulation of Bt-Lon by substrate. In conclusion, structural and biochemical evidence is provided to show that electrostatic interaction in the AAA+ module is important for DNA binding by Lon and allosteric regulation of its enzymatic activities by DNA and substrate.
authors: National Institute of Cancer Research, National Health Research Institutes, Zhunan, Miaoli 35053, Taiwan.
authors: Alan Yueh-Luen
authors: AY
authors: Lee
authors: Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Yu-Da
authors: YD
authors: Chen
authors: Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Yu-Yung
authors: YY
authors: Chang
authors: Institute of Biological Chemistry, Academia Sinica, Taipei 11529, Taiwan.
authors: Yu-Ching
authors: YC
authors: Lin
authors: Genomics Research Center, Academia Sinica, Taipei 11529, Taiwan.
authors: Chi-Fon
authors: CF
authors: Chang
authors: Instrumentation Center, National Taiwan University, Taipei 10617, Taiwan.
authors: Shing-Jong
authors: SJ
authors: Huang
authors: Institute of Biochemical Sciences, National Taiwan University, Taipei 10617, Taiwan.
authors: Shih-Hsiung
authors: SH
authors: Wu
authors: Department of Agricultural Chemistry, National Taiwan University, Taipei 10617, Taiwan.
authors: Chun-Hua
authors: CH
authors: Hsu
mesh_terms: D000494:Allosteric Regulation; D001120:Arginine / Q000737:chemistry* / Q000378:metabolism; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D058675:Brevibacillus / Q000737:chemistry* / Q000201:enzymology; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004269:DNA, Bacterial / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D008958:Models, Molecular; D016296:Mutagenesis; D049070:Protease La / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D055672:Static Electricity; D013816:Thermodynamics
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial; D011994:Recombinant Proteins; D001120:Arginine; D049070:Protease La
keywords: AAA+ proteases; DNA binding; Lon proteases; allosteric regulation