Reconstitution of an active lactose carrier in vivo by simultaneous synthesis of two complementary protein fragments.

Abstract:

:Escherichia coli lactose permease mediates the proton-driven translocation of galactosides across the cytoplasmic membrane. To define regions important for membrane insertion as well as for biological function, we constructed plasmids encoding different portions of the lactose carrier. Among several lacY deletions, two were obtained that encoded mutant proteins with complementary amino acid sequences. The truncated polypeptide Y71/1 (amino acid residues 1 to 71) comprises the first two alpha-helices predicted for the intact protein, and polypeptide delta Y4-69 carries an internal deletion of this region. Regulated coexpression of these lacY-DNA segments governed by separate but identical lacOP control regions resulted in functional complementation with the following characteristics. (i) Simultaneous synthesis of both incomplete proteins restored transport activity in transport-negative cells, measured as accumulation of [14C]lactose. (ii) Under complementing conditions, but not in the absence of the smaller N-terminal protein, specific radiolabeling of the larger polypeptide by N-ethylmaleimide was prevented by substrate. (iii) The presence of the complementing N-terminal polypeptide was also required for the detection of the larger C-terminal protein by antibodies directed against the C terminus of lactose permease, indicating a stabilizing effect contributed by the smaller N-terminal fragment. Thus, coexpression of lacY mutant genes encoding two nonoverlapping portions of the lactose carrier resulted in reconstitution of a two-subunit protein in the cytoplasmic membrane exhibiting biological properties of intact lactose permease.

journal_name

J Bacteriol

journal_title

Journal of bacteriology

authors

Wrubel W,Stochaj U,Sonnewald U,Theres C,Ehring R

doi

10.1128/jb.172.9.5374-5381.1990

subject

Has Abstract

pub_date

1990-09-01 00:00:00

pages

5374-81

issue

9

eissn

0021-9193

issn

1098-5530

journal_volume

172

pub_type

杂志文章
  • Importance of anaerobic superoxide dismutase synthesis in facilitating outgrowth of Escherichia coli upon entry into an aerobic habitat.

    abstract::The manganese-containing isozyme of superoxide dismutase (MnSOD) is synthesized by Escherichia coli only during aerobiosis, in accordance with the fact that superoxide can be formed only in aerobic environments. In contrast, E. coli continues to synthesize the iron-containing isozyme (FeSOD) even in the absence of oxy...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.176.24.7653-7658.1994

    authors: Kargalioglu Y,Imlay JA

    更新日期:1994-12-01 00:00:00

  • ELECTRON TRANSPORT SYSTEM IN VEGETATIVE CELLS AND MICROCYSTS OF MYXOCOCCUS XANTHUS.

    abstract::Dworkin, Martin (University of Minnesota, Minneapolis), and Donald J. Niederpruem. Electron transport system in vegetative cells and microcysts of Myxococcus xanthus. J. Bacteriol. 87:316-322. 1964.-Respiration by intact cells of the fruiting myxobacterium Myxococcus xanthus is cyanide-sensitive and can be demonstrate...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.87.2.316-322.1964

    authors: DWORKIN M,NIEDERPRUEM DJ

    更新日期:1964-02-01 00:00:00

  • Effect of cell cycle position on thermotolerance in Saccharomyces cerevisiae.

    abstract::We showed that the heat killing curve for exponentially growing Saccharomyces cerevisiae was biphasic. This suggests two populations of cells with different thermal killing characteristics. When exponentially growing cells separated into cell cycle-specific fractions via centrifugal elutriation were heat shocked, the ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.169.2.779-784.1987

    authors: Plesset J,Ludwig JR,Cox BS,McLaughlin CS

    更新日期:1987-02-01 00:00:00

  • Iron and Zinc Regulate Expression of a Putative ABC Metal Transporter in Corynebacterium diphtheriae.

    abstract::Corynebacterium diphtheriae, a Gram-positive, aerobic bacterium, is the causative agent of diphtheria and cutaneous infections. While mechanisms required for heme iron acquisition are well known in C. diphtheriae, systems involved in the acquisition of other metals such as zinc and manganese remain poorly characterize...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.00051-18

    authors: Peng ED,Oram DM,Battistel MD,Lyman LR,Freedberg DI,Schmitt MP

    更新日期:2018-04-24 00:00:00

  • Antipain lethality to Escherichia coli: dependence upon cyclic adenosine 3',5'-monophosphate and its receptor protein.

    abstract::Antipain kills Escherichia coli K-12 cells in an exponential manner beginning 1 h after its addition. Mutant strains, delta cya and crp, which are unable to synthesize cyclic adenosine 3',5'-monophosphate (cAMP) and the cAMP receptor protein, respectively, are not affected. Addition of cAMP (5 mM) to antipain-treated ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.139.2.690-693.1979

    authors: Swenson PA

    更新日期:1979-08-01 00:00:00

  • Autoregulation of Escherichia coli purR requires two control sites downstream of the promoter.

    abstract::The expression of Escherichia coli purR, which encodes the pur regulon repressor protein, is autoregulated. Autoregulation at the level of transcription requires two operator sites, designated purRo1 and purRo2 (O1 and O2). Operator O1 is in the region of DNA between the transcription start site and the site for trans...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.172.10.5758-5766.1990

    authors: Rolfes RJ,Zalkin H

    更新日期:1990-10-01 00:00:00

  • Sulfolobus acidocaldarius synthesizes UMP via a standard de novo pathway: results of biochemical-genetic study.

    abstract::A genetic approach was used to establish the route of UMP biosynthesis in Sulfolobus acidocaldarius, a member of the hyperthermophilic division (the Crenarchaeota) of the Archaea domain. Pyrimidine auxotrophs of S. acidocaldarius DG6 were isolated by direct selection and by brute-force methods. Enzymatic assay of extr...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.175.5.1500-1507.1993

    authors: Grogan DW,Gunsalus RP

    更新日期:1993-03-01 00:00:00

  • Germination-defective mutant of Neurospora crassa that responds to siderophores.

    abstract::A conditionally germination-defective mutant of Neurospora crassa has been found to be partially curable by ferricrocin and other siderophores. The mutant conidia rapidly lose their membrane-bound siderophores when suspended in buffer or growth media. Germination is consequently delayed unless large numbers of conidia...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.132.3.1042-1044.1977

    authors: Charlang G,Williams NP

    更新日期:1977-12-01 00:00:00

  • Cell division in Escherichia coli: role of FtsL domains in septal localization, function, and oligomerization.

    abstract::In Escherichia coli, nine essential cell division proteins are known to localize to the division septum. FtsL is a 13-kDa bitopic membrane protein with a short cytoplasmic N-terminal domain, a membrane-spanning segment, and a periplasmic domain that has a repeated heptad motif characteristic of leucine zippers. Here, ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.182.1.116-129.2000

    authors: Ghigo JM,Beckwith J

    更新日期:2000-01-01 00:00:00

  • Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes.

    abstract::Using physical and genetic data, we have demonstrated that Rhizobium meliloti SU47 has a symbiotic megaplasmid, pRmeSU47b, in addition to the previously described nod-nif megaplasmid pRmeSU47a. This plasmid includes four loci involved in exopolysaccharide (exo) synthesis as well as two loci involved in thiamine biosyn...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.167.1.66-72.1986

    authors: Finan TM,Kunkel B,De Vos GF,Signer ER

    更新日期:1986-07-01 00:00:00

  • Growth of flagellar filaments of Escherichia coli is independent of filament length.

    abstract::Bacterial flagellar filaments grow at their distal ends, from flagellin that travels through a central channel ∼2 nm in diameter. The flagellin is extruded from the cytoplasm by a pump powered by a proton motive force (PMF). We measured filament growth in cells near the mid-exponential-phase with flagellin bearing a s...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.06735-11

    authors: Turner L,Stern AS,Berg HC

    更新日期:2012-05-01 00:00:00

  • Expressed genes for plant-type ribulose 1,5-bisphosphate carboxylase/oxygenase in the photosynthetic bacterium Chromatium vinosum, which possesses two complete sets of the genes.

    abstract::Two sets of genes for the large and small subunits of ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) were detected in the photosynthetic purple sulfur bacterium Chromatium vinosum by hybridization analysis with RuBisCO gene probes, cloned by using the lambda Fix vector, and designated rbcL-rbcS and rbcA-rbc...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.171.5.2391-2400.1989

    authors: Viale AM,Kobayashi H,Akazawa T

    更新日期:1989-05-01 00:00:00

  • Three-dimensional structure of the nickel-containing hydrogenase from Thiocapsa roseopersicina.

    abstract::The three-dimensional structure of the nickel-containing hydrogenase from Thiocapsa roseopersicina has been determined at a resolution of 2 nm in the plane and 4 nm in the vertical direction by electron microscopy and computerized image processing on microcrystals of the enzyme. The enzyme forms a large ring-shaped co...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.173.8.2576-2580.1991

    authors: Sherman MB,Orlova EV,Smirnova EA,Hovmöller S,Zorin NA

    更新日期:1991-04-01 00:00:00

  • Gap-filling repair synthesis induced by ultraviolet light in a Bacillus subtilis Uvr- mutant.

    abstract::Deoxyribonucleic acid repair synthesis was studied in one wild-type and two mutant strains of Bacillus subtilis that are defective in excision of pyrimidine dimers. The cells were irradiated with ultraviolet light, and 6-(p-hydroxyphenyl-azo)-uracil was used to block replicative synthesis, allowing only repair synthes...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.139.1.239-246.1979

    authors: Hadden CT

    更新日期:1979-07-01 00:00:00

  • Isolation, sequencing, and mutagenesis of the gene encoding NAD- and glutathione-dependent formaldehyde dehydrogenase (GD-FALDH) from Paracoccus denitrificans, in which GD-FALDH is essential for methylotrophic growth.

    abstract::NAD- and glutathione-dependent formaldehyde dehydrogenase (GD-FALDH) of Paracoccus denitrificans has been purified as a tetramer with a relative molecular mass of 150 kDa. The gene encoding GD-FALDH (flhA) has been isolated, sequenced, and mutated by insertion of a kanamycin resistance gene. The mutant strain is not a...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.177.1.247-251.1995

    authors: Ras J,Van Ophem PW,Reijnders WN,Van Spanning RJ,Duine JA,Stouthamer AH,Harms N

    更新日期:1995-01-01 00:00:00

  • Recombination in the ompA gene but not the omcB gene of Chlamydia contributes to serovar-specific differences in tissue tropism, immune surveillance, and persistence of the organism.

    abstract::Sequences of the major outer membrane protein (MOMP) gene (ompA) and the outer membrane complex B protein gene (omcB) from Chlamydia trachomatis, Chlamydia pneumoniae, and Chlamydia psittaci were analyzed for evidence of intragenic recombination and for linkage equilibrium. The Sawyer runs test, compatibility matrices...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.183.20.5997-6008.2001

    authors: Millman KL,Tavaré S,Dean D

    更新日期:2001-10-01 00:00:00

  • An expression-driven approach to the prediction of carbohydrate transport and utilization regulons in the hyperthermophilic bacterium Thermotoga maritima.

    abstract::Comprehensive analysis of genome-wide expression patterns during growth of the hyperthermophilic bacterium Thermotoga maritima on 14 monosaccharide and polysaccharide substrates was undertaken with the goal of proposing carbohydrate specificities for transport systems and putative transcriptional regulators. Saccharid...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.187.21.7267-7282.2005

    authors: Conners SB,Montero CI,Comfort DA,Shockley KR,Johnson MR,Chhabra SR,Kelly RM

    更新日期:2005-11-01 00:00:00

  • Ribosome synthesis in Rhodopseudomonas capsulata cells growing in continuous and intermittent light.

    abstract::The ribosome content was markedly reduced when illumination (at 28 C) was repeatedly interrupted; an opposite trend was observed with continuous light as the temperature was decreased. ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.97.3.1518-1519.1969

    authors: Din GA,Gest H

    更新日期:1969-03-01 00:00:00

  • Potentiometric titration of the high- and low-potential 4Fe-4S* centers of Azotobacter vinelandii ferredoxin I.

    abstract::The high-potential 4Fe-4S* center ofAzotobacter vinelandii ferredoxin I has been titrated potentiometrically by a reductive procedure. The absorbance decrease at 510 nm accompanying the reduction of the high-potential center titrated with an Em of 320 mV (n = 1). The low-potential 4Fe-4S* center was titrated by using ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.136.2.822-824.1978

    authors: Yoch DC,Carithers RP

    更新日期:1978-11-01 00:00:00

  • Suppression of the defects in rdgB mutants of Escherichia coli K-12 by the cloned purA gene.

    abstract::A gene suppressing the defects of Escherichia coli rdgB (recA-dependent growth) and recA(Ts) rdgB mutants was cloned. The cloned gene suppresses the hyper-Rec phenotype and the enhanced level of SOS functions in rdgB mutants at the nonpermissive temperature. We identified the cloned gene as purA. ...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.173.3.1360-1362.1991

    authors: Clyman J,Cunningham RP

    更新日期:1991-02-01 00:00:00

  • Fatty acid replacements in a fatty acid auxotroph of Escherichia coli.

    abstract::Unsaturated fatty acids having structural features which are different from those of the monoenoic acids normally synthesized by Escherichia coli can serve as growth factors for an auxotroph requiring unsaturated fatty acids. These analogues were incorporated into the phospholipids, as shown by gas-liquid and thin-lay...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.95.5.1658-1665.1968

    authors: Silbert DF,Ruch F,Vagelos PR

    更新日期:1968-05-01 00:00:00

  • Transcriptional regulation and characteristics of a novel N-acetylmuramoyl-L-alanine amidase gene involved in Bacillus thuringiensis mother cell lysis.

    abstract::In Bacillus thuringiensis, a novel N-acetylmuramoyl-L-alanine amidase gene (named cwlB) was detected, and the CwlB protein was purified and characterized. Reverse transcription-PCR (RT-PCR) results indicated that cwlB and an upstream gene (named cwlA) formed one transcriptional unit. 5' rapid amplification of cDNA end...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.00112-13

    authors: Yang J,Peng Q,Chen Z,Deng C,Shu C,Zhang J,Huang D,Song F

    更新日期:2013-06-01 00:00:00

  • Disparate subcellular localization patterns of Pseudomonas aeruginosa Type IV pilus ATPases involved in twitching motility.

    abstract::The opportunistic pathogen Pseudomonas aeruginosa expresses polar type IV pili (TFP), which are responsible for adhesion to various materials and twitching motility on surfaces. Twitching occurs by alternate extension and retraction of TFP, which arise from assembly and disassembly of pilin subunits at the base of the...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.187.3.829-839.2005

    authors: Chiang P,Habash M,Burrows LL

    更新日期:2005-02-01 00:00:00

  • Tetracycline resistance element of pBR322 mediates potassium transport.

    abstract::The tetracycline resistance element of plasmid pBR322 partially complements the potassium transport defect of Escherichia coli K-12 mutants having markedly impaired K+ transport. The plasmid increases K+ transport. The Tn10 element does not result in increased transport, demonstrating that the effect is not general fo...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.160.3.1188-1190.1984

    authors: Dosch DC,Salvacion FF,Epstein W

    更新日期:1984-12-01 00:00:00

  • Allostery of 3-deoxy-D-arabino-heptulosonate 7-phosphate synthetase in Clostridium: another conserved generic characteristic.

    abstract::Enzymological studies were done to characterize the allosteric control of 3-deoxy-d-arabino-heptulosonate 7-phosphate (DAHP) synthetase in three species of Clostridium. Allosteric control was identified as feedback inhibition by phenylalanine and was qualitatively similar for the DAHP synthetases of C. butyricum, C. a...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.111.3.641-648.1972

    authors: Jensen RA,Twarog R

    更新日期:1972-09-01 00:00:00

  • Experimental salmonellosis. VII. Comparison of the immunizing effect of live vaccine and materials extracted from Salmonella enteritidis.

    abstract::Kawakami, Masaya (Gunma University, Maebashi, Japan), Nobutaka Osawa, and Susumu Mitsuhashi. Experimental salmonellosis. VII. Comparison of the immunizing effect of live vaccine and materials extracted from Salmonella enteritidis. J. Bacteriol. 92:1585-1589. 1966.-Attempts were made to isolate the factors of live vacc...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.92.6.1585-1589.1966

    authors: Kawakami M,Osawa N,Mitsuhashi S

    更新日期:1966-12-01 00:00:00

  • Analogs of cyclic AMP as chemoattractants and inhibitors of Dictyostelium chemotaxis.

    abstract::Aggregative amoebae of Dictyostelium discoideum, D. mucoroides, D. purpureum, and D. rosarium react chemotactically to cyclic AMP (cAMP). We measured the chemotactic activity of 14 cAMP analogs and found that these four species have a similar sensitivity to chemical modifications of cAMP; this suggests that the cAMP r...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.149.1.99-105.1982

    authors: Van Haastert PJ,Jastorff B,Pinas JE,Konijn TM

    更新日期:1982-01-01 00:00:00

  • Multicopy derivative of pock-forming plasmid pSA1 in Streptomyces azureus.

    abstract::Streptomyces azureus carried one copy or less of plasmid pSA1, which elicited pocks at 0.1 to 1.0%. Strain PK100 was isolated from the wild-type strain after UV irradiation. PK100 carried approximately 20 to 30 copies of pSA1.1, a derivative of pSA1. Plasmid pSA1.1 elicited pocks at 100% and inhibited spore and thiost...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.168.1.452-454.1986

    authors: Miyoshi YK,Ogata S,Hayashida S

    更新日期:1986-10-01 00:00:00

  • Identification and complementation of frameshift mutations associated with loss of cytadherence in Mycoplasma pneumoniae.

    abstract::Mycoplasma pneumoniae cytadherence is mediated by a specialized, polar attachment organelle. Certain spontaneously arising cytadherence mutants (designated class I) lack HMW2, fail to localize the adhesin protein P1 to the attachment organelle, and exhibit accelerated turnover of proteins HMW1, HMW3, and P65. Insertio...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/JB.181.14.4404-4410.1999

    authors: Fisseha M,Göhlmann HW,Herrmann R,Krause DC

    更新日期:1999-07-01 00:00:00

  • Characterization of endonuclease III (nth) and endonuclease VIII (nei) mutants of Escherichia coli K-12.

    abstract::The nth and nei genes of Escherichia coli affect the production of endonuclease III and endonuclease VIII, respectively, glycosylases/apurinic lyases that attack DNA damaged by oxidizing agents. Here, we provide evidence that oxidative lethal lesions are repaired by both endonuclease III and endonuclease VIII and that...

    journal_title:Journal of bacteriology

    pub_type: 杂志文章

    doi:10.1128/jb.179.11.3783-3785.1997

    authors: Saito Y,Uraki F,Nakajima S,Asaeda A,Ono K,Kubo K,Yamamoto K

    更新日期:1997-06-01 00:00:00