Extensive alanine scanning reveals protein-protein and protein-DNA interaction surfaces in the global regulator FlhD from Escherichia coli.

Abstract:

:FlhD and FlhC are the transcriptional activators of the flagellar regulon. The heterotetrameric complex formed by these two proteins activates the transcription of the class II flagellar genes. The flagellar regulon consists not only of flagellar genes, but also of the chemotactic genes and some receptor proteins. Recently, a connection between the flagellar regulon and some virulence genes has been found in some species. Furthermore, FlhD, but not FlhC, regulates another non-flagellar target. As a first attempt to understand the mechanism of the flagellar transcriptional activation by FlhD and FlhC, the structure of FlhD has been solved. In order to understand the mechanism of the action of FlhD when it regulates the flagellar genes, we conducted site-directed mutagenesis based on its three-dimensional structure. Six interaction surfaces in the FlhD dimer were mapped by alanine scanning mutagenesis. Two of them are surface clusters formed by residues His-2, Asp-28, Arg-35, Phe-34 and Asn-61 located at each side of the dimer core. The other four are located in the flexible arms of the dimer. The residues Ser-82, Arg-83, Val-84, His-91, Thr-92, Ile-94 and Leu-96 are located at this region. All these residues are involved in the FlhD/FlhC interaction with the exception of Ser-82, Arg-83 and Val-84. These three residues affect the DNA-binding ability of the complex. The three-dimensional topology of FlhD and the site-directed mutagenesis results support the hypothesis of FlhC as an allosteric effector that activates FlhD for the recognition of the DNA.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Campos A,Matsumura P

doi

10.1046/j.1365-2958.2001.02248.x

subject

Has Abstract

pub_date

2001-02-01 00:00:00

pages

581-94

issue

3

eissn

0950-382X

issn

1365-2958

pii

mmi2248

journal_volume

39

pub_type

杂志文章
  • Mutualism and pathogenesis in Xenorhabdus and Photorhabdus: two roads to the same destination.

    abstract::Photorhabdus and Xenorhabdus bacteria colonize the intestines of the infective soil-dwelling stage of entomophagous nematodes, Heterorhabditis and Steinernema, respectively. These nematodes infect susceptible insect larvae and release the bacteria into the insect blood. The bacteria kill the insect larvae and convert ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/j.1365-2958.2007.05671.x

    authors: Goodrich-Blair H,Clarke DJ

    更新日期:2007-04-01 00:00:00

  • Experimental genome evolution: large-scale genome rearrangements associated with resistance to replacement of a chromosomal restriction-modification gene complex.

    abstract::Type II restriction enzymes are paired with modification enzymes that protect type II restriction sites from cleavage by methylating them. A plasmid carrying a type II restriction-modification gene complex is not easily replaced by an incompatible plasmid because loss of the former leads to cell death through chromoso...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02436.x

    authors: Handa N,Nakayama Y,Sadykov M,Kobayashi I

    更新日期:2001-05-01 00:00:00

  • Protected environments allow parallel evolution of a bacterial pathogen in a patient subjected to long-term antibiotic therapy.

    abstract::Long-term antibiotic treatment offers a rare opportunity to study the evolution of bacteria within the same individual. The appearance of new variants has been suggested to take place via the selection of enhanced resistance in compartments of the body in which the antibiotic concentration is low. Laboratory models of...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02647.x

    authors: Low AS,MacKenzie FM,Gould IM,Booth IR

    更新日期:2001-11-01 00:00:00

  • The rpf gene of Micrococcus luteus encodes an essential secreted growth factor.

    abstract::Micrococcus luteus secretes a small protein called Rpf, which has autocrine and paracrine signalling functions and is required for the resuscitation of dormant cells. Originally isolated from the supernatant of actively growing cultures, Rpf was also detected on the surface of actively growing bacteria. Most molecules...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2002.03183.x

    authors: Mukamolova GV,Turapov OA,Kazarian K,Telkov M,Kaprelyants AS,Kell DB,Young M

    更新日期:2002-11-01 00:00:00

  • Cloning and characterization of a relA/spoT homologue from Bacillus subtilis.

    abstract::A PCR-amplified DNA fragment of the relA gene from genomic Bacillus subtilis DNA was used to isolate the entire relA/spoT homologue and two adjacent open reading frames (ORFs) from a lambda ZAP Express library. The relA gene, which encodes a protein of 734 amino acid residues (aa), is flanked by an ORF (170aa) that sh...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1997.5511919.x

    authors: Wendrich TM,Marahiel MA

    更新日期:1997-10-01 00:00:00

  • Intramolecular transposition of insertion sequence IS91 results in second-site simple insertions.

    abstract::A series of plasmids carrying an IRL-kan-IRR transposable cassette, in which IRL and IRR are the left- and right-terminal sequences of IS91, have been constructed. These cassettes could be complemented for transposition with similar efficiency when IS91 transposase was provided either in cis or in trans. A total of 87...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1999.01432.x

    authors: Bernales I,Mendiola MV,de la Cruz F

    更新日期:1999-07-01 00:00:00

  • Cloning and expression of a murein hydrolase lipoprotein from Escherichia coli.

    abstract::On the basis of the published N-terminal amino acid sequence of the soluble lytic transglycosylase 35 (Slt35) of Escherichia coli, an open reading frame (ORF) was cloned from the 60.8 min region of the E. coli chromosome. The nucleotide sequence of the ORF, containing a putative lipoprotein-processing site, was shown ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1995.tb02437.x

    authors: Ehlert K,Höltje JV,Templin MF

    更新日期:1995-05-01 00:00:00

  • Spatiotemporal control of FlgZ activity impacts Pseudomonas aeruginosa flagellar motility.

    abstract::The c-di-GMP-binding effector protein FlgZ has been demonstrated to control motility in the opportunistic pathogen Pseudomonas aeruginosa and it was suggested that c-di-GMP-bound FlgZ impedes motility via its interaction with the MotCD stator. To further understand how motility is downregulated in P. aeruginosa and to...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.14236

    authors: Bense S,Bruchmann S,Steffen A,Stradal TEB,Häussler S,Düvel J

    更新日期:2019-06-01 00:00:00

  • Replication strand preference for deletions associated with DNA palindromes.

    abstract::We have isolated and sequenced a set of deletions stimulated by DNA palindromes in Escherichia coli. All of the deletions are asymmetric with respect to the parental sequence and have occurred at short direct repeats. This is consistent with deletion by strand slippage during DNA replication. The orientation of the as...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1998.00831.x

    authors: Pinder DJ,Blake CE,Lindsey JC,Leach DR

    更新日期:1998-05-01 00:00:00

  • Repression of tonB transcription during anaerobic growth requires Fur binding at the promoter and a second factor binding upstream.

    abstract::Although iron is an essential nutrient, its toxicity at high levels necessitates regulated transport. In Gram-negative bacteria a central target for regulation is the TonB protein, an energy transducer that couples the cytoplasmic membrane proton motive force to active transport of (FeIII)-siderophore complexes across...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1994.tb00373.x

    authors: Young GM,Postle K

    更新日期:1994-03-01 00:00:00

  • Septal membrane fusion--a pivotal event in bacterial spore formation?

    abstract::Formation of the asymmetrically located septum divides sporulating bacilli into two distinct cells: the mother cell and the prespore. The rigidifying wall material in the septum is subsequently removed by autolysis. Examination of published electron micrographs indicates that the two septal membranes then fuse to form...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/j.1365-2958.1992.tb01433.x

    authors: Higgins ML,Piggot PJ

    更新日期:1992-09-01 00:00:00

  • Distinct activation mechanisms trigger the trypanocidal activity of DNA damaging prodrugs.

    abstract::Quinone-based compounds have been exploited to treat infectious diseases and cancer, with such chemicals often functioning as inhibitors of key metabolic pathways or as prodrugs. Here, we screened an aziridinyl 1,4-benzoquinone (ABQ) library against the causative agents of trypanosomiasis, and cutaneous leishmaniasis,...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.13767

    authors: Meredith EL,Kumar A,Konno A,Szular J,Alsford S,Seifert K,Horn D,Wilkinson SR

    更新日期:2017-10-01 00:00:00

  • Integration host factor affects expression of two genes at the conjugal transfer origin of plasmid R100.

    abstract::Integration host factor (IHF) binds to two sites near the origin of transfer of the conjugative antibiotic resistance plasmid, R100. DNase I footprinting shows that one site is immediately adjacent to oriT and the gene X promoter, and another is adjacent to the traM promoter. A third site, known only from retardation ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1990.tb00674.x

    authors: Dempsey WB,Fee BE

    更新日期:1990-06-01 00:00:00

  • Pathogen espionage: multiple bacterial adrenergic sensors eavesdrop on host communication systems.

    abstract::The interactions between bacterial pathogens and their eukaryotic hosts are vital in determining the outcome of infections. Bacterial pathogens employ molecular sensors to detect and facilitate adaptation to changes in their niche. The sensing of these extracellular signals enables the pathogen to navigate within mamm...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/mmi.12110

    authors: Karavolos MH,Winzer K,Williams P,Khan CM

    更新日期:2013-02-01 00:00:00

  • Molecular characterization of a surface protein antigen gene from serotype c Streptococcus mutans, implicated in dental caries.

    abstract::The complete nucleotide sequence of the gene for a cell-surface protein antigen (PAc) of Streptococcus mutans MT8148 (serotype c) was determined. The pac gene consisted of 4695 bp and coded for a 170773D protein. The pac gene product contained a putative 38 amino acid signal peptide, resulting in a 166817D mature prot...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1989.tb00215.x

    authors: Okahashi N,Sasakawa C,Yoshikawa M,Hamada S,Koga T

    更新日期:1989-05-01 00:00:00

  • Regulation of polar flagellum genes is mediated by quorum sensing and FlhDC in Burkholderia glumae.

    abstract::The bacterium Burkholderia glumae causes rice grain rot by producing toxoflavin, whose expression is regulated by quorum sensing (QS). We report a major deviation from the current paradigm for the regulation of bacterial polar flagellum genes. The N-octanoyl homoserine lactone (C8-HSL)-deficient mutant of B. glumae is...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2007.05646.x

    authors: Kim J,Kang Y,Choi O,Jeong Y,Jeong JE,Lim JY,Kim M,Moon JS,Suga H,Hwang I

    更新日期:2007-04-01 00:00:00

  • The essential Escherichia coli msbA gene, a multicopy suppressor of null mutations in the htrB gene, is related to the universally conserved family of ATP-dependent translocators.

    abstract::We report the characterization of the msbA gene, isolated as a multicopy suppressor of the HtrB temperature-sensitive phenotype. The msbA gene maps to 20.5 min on the Escherichia coli genetic map and encodes a protein with an estimated molecular mass of 64,460 Da, with the properties of an integral membrane protein. T...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1993.tb01098.x

    authors: Karow M,Georgopoulos C

    更新日期:1993-01-01 00:00:00

  • Characterization of anf genes specific for the alternative nitrogenase and identification of nif genes required for both nitrogenases in Rhodobacter capsulatus.

    abstract::To identify Rhodobacter capsulatus nif genes necessary for the alternative nitrogenase, strains carrying defined mutations in 32 genes and open reading frames of nif region A, B or C were constructed. The ability of these mutants to grow on nitrogen-free medium with molybdenum (Nif phenotype) or in a nifHDK deletion b...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1993.tb01611.x

    authors: Schüddekopf K,Hennecke S,Liese U,Kutsche M,Klipp W

    更新日期:1993-05-01 00:00:00

  • Upregulation of thymidine kinase activity compensates for loss of thymidylate synthase activity in Mycoplasma pneumoniae.

    abstract::Thymidylate, an essential building block of DNA, is synthesized either from deoxyuridylate by thymidylate synthase (TS) or thymidine (dT) by thymidine kinase (TK). Thymidylate kinase (TMPK) phosphorylates dTMP to dTTP. Thymidine phosphorylase (TP) catalyses reversible phosphorolysis of dT. Using transposon mutagenesis...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2010.07298.x

    authors: Wang L,Hames C,Schmidl SR,Stülke J

    更新日期:2010-09-01 00:00:00

  • Characterization of the dTDP-rhamnose biosynthetic genes encoded in the rfb locus of Shigella flexneri.

    abstract::The nucleotide sequence of the proximal half of the rfb region of Shigella flexneri has been determined, and the genes encoding enzymes involved in the biosynthesis of dTDP-rhamnose have been identified. These genes show strong homology to the rfb genes encoding dTDP-rhamnose biosynthesis in Salmonella enterica serova...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1994.tb00308.x

    authors: Macpherson DF,Manning PA,Morona R

    更新日期:1994-01-01 00:00:00

  • Four signalling domains in the hybrid histidine protein kinase RodK of Myxococcus xanthus are required for activity.

    abstract::In prokaryotes, the principal signal transduction systems operating at the level of protein phosphorylation are the two-component systems. A number of hybrid histidine protein kinases in these systems contain several receiver domains, however, the function of these receiver domains is unknown. The RodK kinase in Myxoc...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2006.05118.x

    authors: Rasmussen AA,Wegener-Feldbrügge S,Porter SL,Armitage JP,Søgaard-Andersen L

    更新日期:2006-04-01 00:00:00

  • Lon protease inactivation, or translocation of the lon gene, potentiate bacterial evolution to antibiotic resistance.

    abstract::Previous work demonstrated that selection for Escherichia coli mutants with low antibiotic resistance frequently resulted in co-selection of lon mutations and that lon(-) mutants evolved higher-level resistance faster than a lon(+) strain. Here we show that lon mutation causes a very low multidrug resistance by induci...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.12429

    authors: Nicoloff H,Andersson DI

    更新日期:2013-12-01 00:00:00

  • Cellulosomes: microbial nanomachines that display plasticity in quaternary structure.

    abstract::The assembly of proteins that display complementary activities into supramolecular intra- and extracellular complexes is central to cellular function. One such nanomachine of considerable biological and industrial significance is the plant cell wall degrading apparatus of anaerobic bacteria termed the cellulosome. The...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/j.1365-2958.2007.05640.x

    authors: Gilbert HJ

    更新日期:2007-03-01 00:00:00

  • Functional control of the Candida albicans cell wall by catalytic protein kinase A subunit Tpk1.

    abstract::The cyclic AMP protein kinase A pathway governs numerous biological features of the fungal pathogen Candida albicans. The catalytic protein kinase A subunits, Tpk1 (orf19.4892) and Tpk2 (orf19.2277), have divergent roles, and most studies indicate a more pronounced role for Tpk2. Here we dissect two Tpk1-responsive pr...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2012.08193.x

    authors: Fanning S,Xu W,Beaurepaire C,Suhan JP,Nantel A,Mitchell AP

    更新日期:2012-10-01 00:00:00

  • Small mobile sequences in bacteria display diverse structure/function motifs.

    abstract::Small repeat sequences in bacterial genomes, which represent non-autonomous mobile elements, have close similarities to archaeon and eukaryotic miniature inverted repeat transposable elements. These repeat elements are found in both intergenic and intragenic chromosomal regions, and contain an array of diverse motifs....

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/j.1365-2958.2007.06068.x

    authors: Delihas N

    更新日期:2008-02-01 00:00:00

  • Chaperone-mediated reduction of RepA dimerization is associated with RepA conformational change.

    abstract::RepA, the initiator protein of plasmid P1, binds to multiple sites (iterons) in the origin. The binding normally requires participation of chaperones, DnaJ, DnaK and GrpE. When purified, RepA appears dimeric and is inactive in iteron binding. On reaction with chaperones, a species active in iteron binding is formed an...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1997.5691920.x

    authors: Dibbens JA,Muraiso K,Chattoraj DK

    更新日期:1997-10-01 00:00:00

  • Farnesol and dodecanol effects on the Candida albicans Ras1-cAMP signalling pathway and the regulation of morphogenesis.

    abstract::Candida albicans hypha formation which has been stimulated via the Ras1-cAMP-Efg1 signalling cascade is inhibited by farnesol, a C. albicans autoregulatory factor, and small molecules such as dodecanol. In cultures containing farnesol or dodecanol, hypha formation was restored upon addition of dibutyryl-cAMP. The CAI4...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2007.06013.x

    authors: Davis-Hanna A,Piispanen AE,Stateva LI,Hogan DA

    更新日期:2008-01-01 00:00:00

  • The Saccharomyces cerevisiae Crs5 Metallothionein metal-binding abilities and its role in the response to zinc overload.

    abstract::Crs5 is a Saccharomyces cerevisiae Metallothionein (MT), non-homologous to the paradigmatic Cu-thionein Cup1. Although considered a secondary copper-resistance agent, we show here that it determines survival under zinc overload in a CUP1-null background. Its overexpression prevents the deleterious effects exhibited by...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2006.05510.x

    authors: Pagani A,Villarreal L,Capdevila M,Atrian S

    更新日期:2007-01-01 00:00:00

  • Formation of Rhizobium phaseoli symbiotic plasmids by genetic recombination.

    abstract::We report here the formation of symbiotic plasmids (pSyms), by genetic recombination between rearranged pSyms, which lack symbiotic information, and resistance plasmids carrying parts of different symbiotic plasmids (R's). This recombination was found to occur both between plasmids derived from different Rhizobium pha...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1991.tb00765.x

    authors: Soberón-Chávez G,Nájera R,Espín G,Moreno S

    更新日期:1991-04-01 00:00:00

  • Role of the ribosomal stalk components in the resistance of Aspergillus fumigatus to the sordarin antifungals.

    abstract::Aspergillus fumigatus, an important human nosocomial pathogen, is resistant to sordarin derivatives, a new family of antifungals that inhibit protein synthesis by interaction with the EF-2-ribosomal stalk complex. To explore the role of the A. fumigatus ribosome in the resistance mechanism, the fungal stalk proteins w...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2002.02736.x

    authors: Santos C,Ballesta JP

    更新日期:2002-01-01 00:00:00