Characterization and modelling of VanT: a novel, membrane-bound, serine racemase from vancomycin-resistant Enterococcus gallinarum BM4174.

Abstract:

:Sequence determination of a region downstream from the vanXYc gene in Enterococcus gallinarum BM4174 revealed an open reading frame, designated vanT, that encodes a 698-amino-acid polypeptide with an amino-terminal domain containing 10 predicted transmembrane segments. The protein contained a highly conserved pyridoxal phosphate attachment site in the C-terminal domain, typical of alanine racemases. The protein was overexpressed in Escherichia coli, and serine racemase activity was detected in the membrane but not in the cytoplasmic fraction after centrifugation of sonicated cells, whereas alanine racemase activity was located almost exclusively in the cytoplasm. When the protein was overexpressed as a polypeptide lacking the predicted transmembrane domain, serine racemase activity was detected in the cytoplasm. The serine racemase activity was partially (64%) inhibited by D-cycloserine, whereas host alanine racemase activity was almost totally inhibited (97%). Serine racemase activity was also detected in membrane preparations of constitutively vancomycin-resistant E. gallinarum BM4174 but not in BM4175, in which insertional inactivation of the vanC-1 D-Ala:D-Ser ligase gene probably had a polar effect on expression of the vanXYc and vanT genes. Comparative modelling of the deduced C-terminal domain was based on the alignment of VanT with the Air alanine racemase from Bacillus stearothermophilus. The model revealed that almost all critical amino acids in the active site of Air were conserved in VanT, indicating that the C-terminal domain of VanT is likely to adopt a three-dimensional structure similar to that of Air and that the protein could exist as a dimer. These results indicate that the source of D-serine for peptidoglycan synthesis in vancomycin-resistant enterococci expressing the VanC phenotype involves racemization of L- to D-serine by a membrane-bound serine racemase.

journal_name

Mol Microbiol

journal_title

Molecular microbiology

authors

Arias CA,Martín-Martinez M,Blundell TL,Arthur M,Courvalin P,Reynolds PE

doi

10.1046/j.1365-2958.1999.01294.x

subject

Has Abstract

pub_date

1999-03-01 00:00:00

pages

1653-64

issue

6

eissn

0950-382X

issn

1365-2958

journal_volume

31

pub_type

杂志文章
  • A fresh view of the cell biology of copper in enterobacteria.

    abstract::Copper ions are essential but also very toxic. Copper resistance in bacteria is based on export of the toxic ion, oxidation from Cu(I) to Cu(II), and sequestration by copper-binding metal chaperones, which deliver copper ions to efflux systems or metal-binding sites of copper-requiring proteins. In their publication i...

    journal_title:Molecular microbiology

    pub_type: 评论,杂志文章

    doi:10.1111/mmi.12123

    authors: Nies DH,Herzberg M

    更新日期:2013-02-01 00:00:00

  • A MAP kinase encoded by the ubc3 gene of Ustilago maydis is required for filamentous growth and full virulence.

    abstract::Ustilago maydis, the causal agent of corn smut disease, displays dimorphic growth in which it alternates between a budding haploid saprophyte and a filamentous dikaryotic pathogen. We are interested in identifying the genetic determinants of filamentous growth and pathogenicity in U. maydis. To do this, we have taken ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1999.01610.x

    authors: Mayorga ME,Gold SE

    更新日期:1999-11-01 00:00:00

  • The link between 20S proteasome activity and post-replication DNA repair in Saccharomyces cerevisiae.

    abstract::We have shown previously that deletion of the Saccharomyces cerevisiae UMP1 gene encoding the 20S proteasome maturase causes sensitivity to UV radiation. In the current report, we have extended this finding to show that mutations specifically compromising chymotrypsin-like or trypsin-like activity of 20S proteasome pe...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2003.03635.x

    authors: Podlaska A,McIntyre J,Skoneczna A,Sledziewska-Gojska E

    更新日期:2003-09-01 00:00:00

  • An increase in expression of a Mycobacterium tuberculosis mycolyl transferase gene (fbpB) occurs early after infection of human monocytes.

    abstract::Changes in the mRNA levels of two Mycobacterium tuberculosis genes (fbpB known as antigen 85B, and hspX known as Acr) were studied in infected human monocytes. Antigen 85B is an enzyme involved in cell wall biosynthesis and is also a major target of the immune response. Acr is a stress protein believed to be involved ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02280.x

    authors: Wilkinson RJ,DesJardin LE,Islam N,Gibson BM,Kanost RA,Wilkinson KA,Poelman D,Eisenach KD,Toossi Z

    更新日期:2001-02-01 00:00:00

  • The Rcs phosphorelay system is specific to enteric pathogens/commensals and activates ydeI, a gene important for persistent Salmonella infection of mice.

    abstract::Bacteria utilize phosphorelay systems to respond to environmental or intracellular stimuli. Salmonella enterica encodes a four-step phosphorelay system that involves two sensor kinase proteins, RcsC and RcsD, and a response regulator, RcsB. The physiological stimulus for Rcs phosphorelay activation is unknown; however...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2006.05420.x

    authors: Erickson KD,Detweiler CS

    更新日期:2006-11-01 00:00:00

  • Disruption of msl3 abolishes the synthesis of mycolipanoic and mycolipenic acids required for polyacyltrehalose synthesis in Mycobacterium tuberculosis H37Rv and causes cell aggregation.

    abstract::Cell wall lipids of Mycobacterium tuberculosis containing multiple methylbranched fatty acids play critical roles in pathogenesis and thus offer targets for new antimycobacterial drugs. Mycocerosicacid synthase gene (mas) encodes the enzyme that produces one class of such acids. Seven mas-like genes (msls) were identi...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2002.03119.x

    authors: Dubey VS,Sirakova TD,Kolattukudy PE

    更新日期:2002-09-01 00:00:00

  • Cytotoxic activity of a recombinant chimaeric protein between Pseudomonas aeruginosa exotoxin A and Corynebacterium diphtheriae diphtheria toxin.

    abstract::A segment of the exotoxin A gene of Pseudomonas aeruginosa, coding for the N-terminal end of domain I and domain II of the toxin (ETA), was genetically fused to the diphtheria toxin gene of Corynebacterium diphtheriae, coding for the N-terminal end of A fragment of diphtheria toxin (DT). The resulting hybrid protein (...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1992.tb00849.x

    authors: Guidi-Rontani C

    更新日期:1992-05-01 00:00:00

  • Effects of DksA and ClpP protease on sigma S production and virulence in Salmonella typhimurium.

    abstract::Salmonella typhimurium responds to a variety of environmental stresses by accumulating the alternative sigma factor sigmaS. The repertoire of sigmaS -dependent genes that are subsequently expressed confers tolerance to a variety of potentially lethal conditions including low pH and stationary phase. The mechanism(s) r...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.1999.01581.x

    authors: Webb C,Moreno M,Wilmes-Riesenberg M,Curtiss R 3rd,Foster JW

    更新日期:1999-10-01 00:00:00

  • Multifunctional analysis of Chlamydia-specific genes in a yeast expression system.

    abstract::Our understanding of how obligate intracellular pathogens co-opt eukaryotic cellular functions has been limited by their intractability to genetic manipulation and by the abundance of pathogen-specific genes with no known functional homologues. In this report we describe a gene expression system to characterize protei...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2006.05074.x

    authors: Sisko JL,Spaeth K,Kumar Y,Valdivia RH

    更新日期:2006-04-01 00:00:00

  • Control of FimY translation and type 1 fimbrial production by the arginine tRNA encoded by fimU in Salmonella enterica serovar Typhimurium.

    abstract::Expression of type 1 fimbriae in Salmonella enterica serovar Typhimurium undergoes phase variation or alteration between a fimbriate and a non-fimbriate phenotype. This variation is known to be dependent upon environmental conditions in vitro and is thought to be a complex process involving regulation by a number of p...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02430.x

    authors: Tinker JK,Clegg S

    更新日期:2001-05-01 00:00:00

  • Requirement of essential Pbp2x and GpsB for septal ring closure in Streptococcus pneumoniae D39.

    abstract::Bacterial cell shapes are manifestations of programs carried out by multi-protein machines that synthesize and remodel the resilient peptidoglycan (PG) mesh and other polymers surrounding cells. GpsB protein is conserved in low-GC Gram-positive bacteria and is not essential in rod-shaped Bacillus subtilis, where it pl...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.12408

    authors: Land AD,Tsui HC,Kocaoglu O,Vella SA,Shaw SL,Keen SK,Sham LT,Carlson EE,Winkler ME

    更新日期:2013-12-01 00:00:00

  • Temperature effect on inclusion body formation and stress response in the periplasm of Escherichia coli.

    abstract::We previously characterized a defective-folding mutant of maltose-binding protein of Escherichia coli, MalE31, which formed periplasmic inclusion bodies. Here, we show that MalE31 aggregation does not affect bacterial growth at 30 degrees C but is lethal at 37 degrees C. Surprisingly, under mild heat shock conditions ...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2003.03785.x

    authors: Hunke S,Betton JM

    更新日期:2003-12-01 00:00:00

  • Use of rare-earth elements in the phyllosphere colonizer Methylobacterium extorquens PA1.

    abstract::Until recently, rare-earth elements (REEs) had been thought to be biologically inactive. This view changed with the discovery of the methanol dehydrogenase XoxF that strictly relies on REEs for its activity. Some methylotrophs only contain xoxF, while others, including the model phyllosphere colonizer Methylobacterium...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.14208

    authors: Ochsner AM,Hemmerle L,Vonderach T,Nüssli R,Bortfeld-Miller M,Hattendorf B,Vorholt JA

    更新日期:2019-05-01 00:00:00

  • The unusual mycobacterial chaperonins: evidence for in vivo oligomerization and specialization of function.

    abstract::The pathogen Mycobacterium tuberculosis expresses two chaperonins, one (Cpn60.1) dispensable and one (Cpn60.2) essential. These proteins have been reported not to form oligomers despite the fact that oligomerization of chaperonins is regarded as essential for their function. We show here that the Cpn60.2 homologue fro...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2012.08150.x

    authors: Fan M,Rao T,Zacco E,Ahmed MT,Shukla A,Ojha A,Freeke J,Robinson CV,Benesch JL,Lund PA

    更新日期:2012-09-01 00:00:00

  • Involvement of surface polysaccharides in the organic acid resistance of Shiga Toxin-producing Escherichia coli O157:H7.

    abstract::In general, wild Escherichia coli strains can grow effectively under moderately acidic organic acid-rich conditions. We found that the Shiga Toxin-producing E. coli (STEC) O157:H7 NGY9 grows more quickly than a K-12 strain in Luria-Bertani (LB)-2-morpholinoethanesulphonic acid (MES) broth supplemented with acetic acid...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2002.02768.x

    authors: Barua S,Yamashino T,Hasegawa T,Yokoyama K,Torii K,Ohta M

    更新日期:2002-02-01 00:00:00

  • Signalling substitutions in the periplasmic domain of chemoreceptor Trg induce or reduce helical sliding in the transmembrane domain.

    abstract::We used in vivo oxidative cross-linking of engineered cysteine pairs to assess conformational changes in the four-helix transmembrane domain of chemoreceptor Trg. Extending previous work, we searched for and found a fourth cross-linking pair that spanned the intrasubunit interface between transmembrane helix 1 (TM1) a...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02446.x

    authors: Beel BD,Hazelbauer GL

    更新日期:2001-05-01 00:00:00

  • The DivJ, CbrA and PleC system controls DivK phosphorylation and symbiosis in Sinorhizobium meliloti.

    abstract::Sinorhizobium meliloti is a soil bacterium that invades the root nodules it induces on Medicago sativa, whereupon it undergoes an alteration of its cell cycle and differentiates into nitrogen-fixing, elongated and polyploid bacteroid with higher membrane permeability. In Caulobacter crescentus, a related alphaproteoba...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.12347

    authors: Pini F,Frage B,Ferri L,De Nisco NJ,Mohapatra SS,Taddei L,Fioravanti A,Dewitte F,Galardini M,Brilli M,Villeret V,Bazzicalupo M,Mengoni A,Walker GC,Becker A,Biondi EG

    更新日期:2013-10-01 00:00:00

  • Cytoplasmic membrane protonmotive force energizes periplasmic interactions between ExbD and TonB.

    abstract::The TonB system of Escherichia coli (TonB/ExbB/ExbD) transduces the protonmotive force (pmf) of the cytoplasmic membrane to drive active transport by high-affinity outer membrane transporters. In this study, chromosomally encoded ExbD formed formaldehyde-linked complexes with TonB, ExbB and itself (homodimers) in vivo...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2009.06785.x

    authors: Ollis AA,Manning M,Held KG,Postle K

    更新日期:2009-08-01 00:00:00

  • Mycobacterium smegmatis RNA polymerase: DNA supercoiling, action of rifampicin and mechanism of rifampicin resistance.

    abstract::We have isolated RNA polymerase from Mycobacterium smegmatis and established conditions for specific transcription initiation in vitro. The M. smegmatis enzyme has a strong dependence on supercoiling of the DNA substrate for transcription from mycobacterial promoters. We also show that RNA polymerase is the target for...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1993.tb01572.x

    authors: Levin ME,Hatfull GF

    更新日期:1993-04-01 00:00:00

  • Analysis of the replicon region and identification of an rRNA operon on pBM400 of Bacillus megaterium QM B1551.

    abstract::An 18 633 bp region containing the replicon from the approximately 53 kb pBM400 plasmid of Bacillus megaterium QM B1551 has been sequenced and characterized. This region contained a complete rRNA operon plus 10 other potential open reading frames (ORFs). The replicon consisted of an upstream promoter and three contigu...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2001.02292.x

    authors: Kunnimalaiyaan M,Stevenson DM,Zhou Y,Vary PS

    更新日期:2001-02-01 00:00:00

  • The bacterial porin superfamily: sequence alignment and structure prediction.

    abstract::The porins of Gram-negative bacteria are responsible for the 'molecular sieve' properties of the outer membrane. They form large water-filled channels which allow the diffusion of hydrophilic molecules into the periplasmic space. Owing to the strong hydrophilicity of their amino acid sequence and the nature of their s...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1991.tb02145.x

    authors: Jeanteur D,Lakey JH,Pattus F

    更新日期:1991-09-01 00:00:00

  • Isolation of arginine auxotrophs, cloning by mutant complementation, and sequence analysis of the argC gene from the cyanobacterium Anabaena species PCC 7120.

    abstract::Arginine auxotrophs of the dinitrogen-fixing cyanobacterium Anabaena species strain PCC 7120 were isolated after ultraviolet light mutagenesis and penicillin enrichment. Two of these auxotrophs were complemented by a cosmid gene library of the wild-type strain established in Escherichia coli that was transferred en ma...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1992.tb01381.x

    authors: Floriano B,Herrero A,Flores E

    更新日期:1992-08-01 00:00:00

  • Leishmania mexicana can utilize amino acids as major carbon sources in macrophages but not in animal models.

    abstract::Leishmania parasites target macrophages in their mammalian hosts and proliferate within the mature phagolysosome compartment of these cells. Intracellular amastigote stages are dependent on sugars as a major carbon source in vivo, but retain the capacity to utilize other carbon sources. To investigate whether amastigo...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.13923

    authors: Saunders EC,Naderer T,Chambers J,Landfear SM,McConville MJ

    更新日期:2018-04-01 00:00:00

  • Competitive activation of the Escherichia coli argO gene coding for an arginine exporter by the transcriptional regulators Lrp and ArgP.

    abstract::In vivo and in vitro analyses indicate that transcription of the argO gene coding for an arginine exporter is regulated by the global transcriptional regulator Lrp, an effect that went by unnoticed in previous genome-scale screenings of the Lrp regulatory network in Escherichia coli. Lrp activates the argO promoter fo...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2009.06950.x

    authors: Peeters E,Nguyen Le Minh P,Foulquié-Moreno M,Charlier D

    更新日期:2009-12-01 00:00:00

  • Regulation of the KstR2 regulon of Mycobacterium tuberculosis by a cholesterol catabolite.

    abstract::Cholesterol catabolism is widespread in actinobacteria and is critical for Mycobacterium tuberculosis (Mtb) virulence. Catabolism of steroid nucleus rings C and D is poorly understood: it is initiated by the CoA thioesterification of 3aα-H-4α(3'-propanoate)-7aβ-methylhexahydro-1,5-indanedione (HIP) by FadD3, whose gen...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/mmi.12340

    authors: Casabon I,Zhu SH,Otani H,Liu J,Mohn WW,Eltis LD

    更新日期:2013-09-01 00:00:00

  • Genetic and functional analysis of the phosphorylcholine moiety of commensal Neisseria lipopolysaccharide.

    abstract::Phosphorylcholine (ChoP) is a common surface feature of many mucosal organisms, including Neisseria spp., in which it is present exclusively on pili of pathogenic Neisseria and on the lipopolysaccharide (LPS) of commensal Neisseria (Cn). Its presence in Cn has been confirmed by nuclear magnetic resonance. It appears t...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1046/j.1365-2958.2002.02755.x

    authors: Serino L,Virji M

    更新日期:2002-01-01 00:00:00

  • A mixed double negative feedback loop between the sRNA MicF and the global regulator Lrp.

    abstract::Roughly 10% of all genes in Escherichia coli are controlled by the global transcription factor Lrp, which responds to nutrient availability. Bioinformatically, we identified lrp as one of several putative targets for the sRNA MicF, which is transcriptionally downregulated by Lrp. Deleting micF results in higher Lrp le...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2012.07994.x

    authors: Holmqvist E,Unoson C,Reimegård J,Wagner EG

    更新日期:2012-05-01 00:00:00

  • Control of Cre recombination by regulatory elements from Xer recombination systems.

    abstract::Site-specific recombination by the Cre recombinase takes place at a simple DNA site (loxP), requires no additional proteins and gives topologically simple recombination products. In contrast, cer and psi sites for Xer recombination contain approximately 150 bp of accessory sequences, require accessory proteins PepA, A...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.2003.03962.x

    authors: Gourlay SC,Colloms SD

    更新日期:2004-04-01 00:00:00

  • Regulatory noise in prokaryotic promoters: how bacteria learn to respond to novel environmental signals.

    abstract::Various features of the regulation of pathways for biodegradation of recalcitrant compounds by Pseudomonas provide insights into the mechanisms by which operons evolve to acquire conditionally active promoters that permit the corresponding genes to be transcribed only when required. The "regulatory noise hypothesis' p...

    journal_title:Molecular microbiology

    pub_type: 杂志文章,评审

    doi:10.1111/j.1365-2958.1996.tb02463.x

    authors: de Lorenzo V,Pérez-Martín J

    更新日期:1996-03-01 00:00:00

  • The transcriptional organization of the Bacillus subtilis 168 chromosome region between the spoVAF and serA genetic loci.

    abstract::The genetic organization of the spoVAF-serA area of the Bacillus subtilis chromosome and its putative transcription map have been derived from analysis of the nucleotide sequence. In order to confirm this transcription map as regards size of transcripts and to determine growth conditions for their appearance, we under...

    journal_title:Molecular microbiology

    pub_type: 杂志文章

    doi:10.1111/j.1365-2958.1993.tb02671.x

    authors: Azevedo V,Sorokin A,Ehrlich SD,Serror P

    更新日期:1993-10-01 00:00:00