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Biomedical subjects

M M Gibson

Publications and source records attributed to M M Gibson.

11 recordsLinked to original sources

Peptide transport and chemotaxis in Escherichia coli and Salmonella typhimurium: characterization of the dipeptide permease (Dpp) and the dipeptide-binding protein.

The dipeptide permease (Dpp) is one of three genetically distinct peptide-transport systems in enteric bacteria. Dpp also plays a role in chemotaxis towards peptides. We have devised three selections for dpp mutations based on resistance to toxic peptides (bacilysin, valine-containing peptides, and bialaphos). All dpp mutations mapped to a single chromosomal locus between 77 and 78 min in Salmonella typhimurium and at 79.2 min in Escherichia coli. Expression of dpp was constitutive in both species but the absolute level of expression varied widely between strains. At least in part this difference in expression levels is determined by cis-acting sequences. The dpp locus of E. coli was cloned. The first gene in the operon, dppA, encodes a periplasmic dipeptide-binding protein (DBP) required for dipeptide transport and chemotaxis. Downstream of dppA are other genes required for transport but not for chemotaxis. The dipeptide-binding protein was found to share 26.5% sequence identity with the periplasmic oligopeptide-binding protein OppA.

Amino Acid Sequence↗

Magnesium transport in Salmonella typhimurium: the influence of new mutations conferring Co2+ resistance on the CorA Mg2+ transport system.

The CorA Mg2+ transport system of Salmonella typhimurium mediates both influx and efflux of Mg2+. Mutations at the corA locus (83.5 min) confer resistance to Co2+. Using transposon mutagenesis, three additional Co2+ resistance loci (corB, corC, and corD) were found and mapped to 55, 15, and 3min, respectively, on the S. typhimurium chromosome. No mutations corresponding to the reported corB locus at 95 min in Escherichia coli were obtained. The corB, corC, and corD mutations confer levels of Co2+ resistance intermediate between those of the wild-type and corA mutations. Isogenic strains were constructed containing combinations of transposon insertion mutations in each of the three Co(2+)-resistance loci to assess their influence on the CorA Mg2+ transport system. The Vmax and Km values for 28Mg2+ or for 57Co2+ and 63Ni2+ influx, analogues of Mg2+ transported by the CorA system, were changed less than twofold compared with the wild-type values, regardless of the mutation(s) present. However, while efflux of 28Mg2+ through the CorA system was decreased threefold in strains carrying one or two mutant alleles among corB, corC, or corD, efflux was completely abolished in either a corA or a corBCD strain. Thus, although the corA gene product is necessary and sufficient to mediate Mg2+ influx, Mg2+ efflux requires the presence of a wild-type allele of at least one of the corB, corC or corD loci.

Bacterial Proteins↗

OmpR and EnvZ are pleiotropic regulatory proteins: positive regulation of the tripeptide permease (tppB) of Salmonella typhimurium.

The tppB locus of Salmonella typhimurium encodes the anaerobically-induced tripeptide permease. We have demonstrated that expression of tppB requires the function of the ompR and envZ gene products, originally identified as positive regulatory proteins required for the osmotic regulation of porin expression. Significantly, tppB expression is not osmotically regulated. We have also identified three additional genes whose expression depends on OmpR. Thus OmpR and EnvZ serve a more general regulatory role than has previously been supposed. This study provides the first detailed genetic analysis of the ompB locus of S. typhimurium.

Amino Acid Sequence↗

Wiener kernel analysis of responses from anteroventral cochlear nucleus neurons.

Responses to pseudo-random Gaussian white noise, tones and clics were recorded from neurons in the anteroventral cochlear nucleus (AVCN) of barbiturate anesthetized cats. The responses to white noise were used to calculate estimates of the zero-, first- and second-order Wiener kernels for these neurons. The Wiener kernels did contain useful information on the fundamental, DC and second harmonic components of the responses of AVCN neurons to tones, clicks and noise. However, they generally did not provide predictions of the difference tone distortion products found in the peripheral auditory system. Overall, the addition of the second kernel improved a prediction based on the zero- and first-order kernels, but not by very much. If the estimates of the Wiener kernels were not very good, then a second-order prediction could be worse than a first-order one. To produce good estimates of the Wiener kernels, many repetitions of very long Gaussian white noise stimuli are necessary. Therefore the technique does not permit rapid data collection. Further, exposure to long duration high intensity noise can result in acoustic trauma. This damage effects the mechanism that generates the difference tone distortion products, and it can also affect the tuning of the auditory neurons. Thus Wiener's nonlinear system identification theory has only limited usefulness in the analysis of the peripheral auditory system.

Acoustic Stimulation↗

Genetic characterization and molecular cloning of the tripeptide permease (tpp) genes of Salmonella typhimurium.

Of the three bacterial peptide transport systems only one, the oligopeptide permease, has been characterized in any detail. We have now isolated Salmonella typhimurium mutants deficient in a second transport system, the tripeptide permease (Tpp), using the toxic peptide alafosfalin. Alafosfalin resistance mutations map at three loci, the gene encoding peptidase A (pepA) and two transport-defective loci, tppA and tppB. Locus tppA has been mapped to 74 min on the S. typhimurium chromosome, cotransducible with aroB, and is a positive regulator of tppB. Locus tppB maps at 27 min in the cotransduction gap between purB and pyrF. We cloned tppB, the structural locus for the tripeptide permease. Two simple methods are described for mapping the location of cloned DNA fragments on the chromosome of S. typhimurium.

Alanine↗

Responses of single units in the cat cochlear nucleus to sinusoidal amplitude modulation of tones and noise: linearity and relation to speech perception.

Responses to the sinusoidal modulation envelopes of amplitude-modulated tonend noise carriers were recorded from single units in the cochlear nucleus of the cat. The unit discharges were synchronized to the peaks of the modulation envelope. Population-averaged firing rate were independent of the modulation index of the stimulus. Temporal firing patterns, as represented by the shapes and magnitudes of modulation-cycle histograms, were strongly dependent on stimulus intesity and modulation index. Nonlinear nonsinusoidal responses to sinusoidal modulation envelopes were observed, but only at high values of sound intensity. These and other results are discussed in the context of psychological studies concerning the perception of speech information.

Acoustic Stimulation↗