{"refrec":{"BRefID":281040,"RR":"<b>Xu, Y.; Sun, Y.; Huysveld, N.; Gigot, D.; Glansdorff, N.; Charlier, D.</b> (2003). Regulation of arginine biosynthesis in the psychropiezophilic bacterium <i>Moritella profunda</i>: <i>in vivo</i> repressibility and <i>in vitro</i> repressor-operator contact probing. <i>J. Mol. Biol. 326(2)</i>: 353-369. <a href=\"https://dx.doi.org/10.1016/S0022-2836(02)01375-X\" target=\"_blank\">https://dx.doi.org/10.1016/S0022-2836(02)01375-X</a>","BEntID":273059,"PublicFlag":1,"CheckedFlag":1,"wosflag":1,"vabbflag":1,"RefStringPartII":". <i>J. Mol. Biol. 326(2)</i>: 353-369. <a href=\"https://dx.doi.org/10.1016/S0022-2836(02)01375-X\" target=\"_blank\">https://dx.doi.org/10.1016/S0022-2836(02)01375-X</a>","DocTypID":8,"DocType":"Journal article","MarineFlag":1,"FreshFlag":0,"BrackishFlag":0,"TerrestrialFlag":0,"Authorstring":"Xu, Y.; Sun, Y.; Huysveld, N.; Gigot, D.; Glansdorff, N.; Charlier, D.","OrigTitleTranslFlag":0,"Authorstringtrunc":"Xu, Y. <i>et al.</i>","Englishabstract":"We report the cloning of the arginine repressor gene from the psychropiezophilic Gram-negative bacterium <i>Moritella profunda</i>, the purification of its product (ArgR<sub>Mp</sub>), the identification of the operator in the bipolar <i>argECBFGH</i>(<i>A</i>) operon, <i>in vivo</i> repressibility studies, and an <i>in vitro</i> analysis of the repressor–operator interaction, including binding to mutant and heterologous arginine operators. The ArgR<sub>Mp</sub> subunit shows about 70% amino acid sequence identity with <i>Escherichia coli</i> ArgR (ArgR<sub>Ec</sub>). Binding of purified hexameric ArgR<sub>Mp</sub> to the control region of the divergent operon proved to be arginine-dependent, sequence-specific, and significantly more sensitive to heat than complex formation with ArgR<sub>Ec</sub>. ArgR<sub>Mp</sub> binds <i>E. coli</i> arginine operators very efficiently, but hardly recognizes the operator from <i>Bacillus stearothermophilus</i> or <i>Thermotoga maritima</i>. ArgR<sub>Mp</sub> binds to a single site overlapping the −35 element of <i>argC</i><sub>P</sub>, but not <i>argE</i><sub>P</sub>. Therefore, the arrangement of promoter and operator sites in the bipolar <i>argECBFGH</i>(<i>A</i>) operon of <i>M. profunda</i> is very different from the organization of control elements in the bipolar <i>argECBH</i> operon of <i>E. coli</i>, where both promoters overlap the common operator and are equally repressible. We demonstrate that <i>M. profunda</i><i>argC</i><sub><i>P</i></sub> is about 44-fold repressible, whereas <i>argE</i><sub>P</sub> is fully constitutive. A high-resolution contact map of the ArgR<sub>Mp</sub>–operator interaction was established by enzymatic and chemical footprinting, missing contact and base-specific premodification binding interference studies. The results indicate that the <i>argC</i> operator consists of two ARG box-like sequences (18 bp imperfect palindromes) separated by 3 bp. ArgR<sub>Mp</sub> binds to one face of the DNA helix and establishes contacts with two major groove segments and the intervening minor groove of each ARG box, whereas the minor groove segment facing the repressor at the center of the operator remains largely uncontacted. This pattern is reminiscent of complex formation with the repressors of <i>E. coli</i> and <i>B. stearothermophilus</i>, and suggests that each ARG box is contacted by two ArgR subunits belonging to opposite trimers. Moreover, the premodification interference patterns and mutant studies clearly indicate that the inner, center proximal halves of each ARG box in the <i>M. profunda</i><i>argC</i> operator are more important for complex formation and repression than the outermost halves. A close inspection of sequence conservation and of single base-pair O<sup>c</sup>-type mutations indicate that the same conclusion can be generalized to <i>E. coli</i> operators.","AbstractOtherLang":null,"BibLvlCode":"AS","StandardTitle":"Regulation of arginine biosynthesis in the psychropiezophilic bacterium <i>Moritella profunda</i>: <i>in vivo</i> repressibility and <i>in vitro</i> repressor-operator contact probing","OrigTitleLangCode":"en","OrigTitleLangCodeExtended":"eng","OrigTitleLangID":15,"DateLastModified":{"date":"2026-06-15 01:32:57.379041","timezone_type":1,"timezone":"+02:00"},"UserAccessRight":null,"UserAccID":null,"AuthorKeywords":"Moritella profunda; psychrophiles; arginine metabolism; transcriptionregulation; protein-DNA contacts","OtherDescriptors":null,"Notes":null,"AnaPub":2003,"MonPub":null,"DateUpdate":"2022-07-12","DateCreate":"2016-10-09","SecASFANote":null,"ConfID":null,"PeerRev":1,"VlizCoreFlag":1,"WoScode":"WOS:000181068900002","VABBcode":null,"OpenAcc":0,"DOI":"10.1016/S0022-2836(02)01375-X"},"refs":null,"anarec":{"AnaID":281040,"PubliDate":2003,"Pagination":"353-369","XtraPublOfAnaID":null,"ISBN":null,"Volume":"326","Issue":"2","BRefMon":null,"BRefMonRR":null,"BRefXtra":null,"BRefXtraRR":null,"SerBRefID":43083,"SerRR":"Journal of Molecular Biology. 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