Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “transducer selectivity”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 19 recordsLinked to original sources
PubMed2026

Structural basis of β-arrestin coupling and transducer selectivity in PAC1R.

The pituitary adenylate cyclase-activating polypeptide receptor (PAC1R) is a class B G protein-coupled receptor (GPCR) that engages both G proteins and β-arrestins to mediate diverse signaling responses, yet how PAC1R adopts distinct intracellular conformations to achieve this transducer selectivity remains poorly understood. Here, we report the cryo-electron microscopy structure of PAC1R in complex with β-arrestin 1 (βarr1), revealing a core-engaged conformation. Comparison with the Gs-bound PAC1R structure shows that βarr1 engagement is associated with remodeling of the intracellular transmembrane bundle, including TM5 reorientation and inward movement of TM6, resulting in a receptor core geometry distinct from that of the G protein-bound state. Comparison with the βarr1-bound parathyroid hormone receptor 1 (PTH1R) structure further reveals both conserved and receptor-specific features of βarr1 engagement. Although outward displacement of the TM5 cytoplasmic end is observed in both PAC1R-βarr1 and PTH1R-βarr1 complexes, its specific direction and the resulting TM5-TM6 rearrangements differ between receptors, correlating with distinct βarr1 finger loop orientations within the receptor core. Together, these findings suggest that β-arrestin core engagement by class B GPCRs is accompanied by receptor-specific intracellular remodeling that may contribute to transducer selectivity in PAC1R.

Class B GPCR↗
PubMed1979

A technique for measuring frequency response of pressure, volume, and flow transducers.

A device and methodology is presented for testing the frequency response of pressure, volume, or flow transducers. Also reported are responses of selected transducers of all three types over the range of 2--120 Hz. Several pressure transducers tested had good frequency response when connected to the test system with a minimum of interconnecting fittings; others did not. Use of additional connectors degraded the response as did the addition of air-filled catheters. The frequency response of the pneumotachometers tested were influenced largely by the response characteristics of the associated pressure transducer and interconnecting fittings. These results emphasize the need to test the response characteristics of any transducer with specific connectors and fittings that are to be used to make the actual measurements of pressure, volume, or flow.

Mathematics↗
PubMed1979

Differential diagnosis of hypoechoic and anechoic masses with gray scale sonography: new observations.

With the technological advances in gray scale sonography that have permitted the use of higher-frequency transducers and expansion of the acoustic dynamic range, increasing problems in differentiating solid masses and fluid-filled masses have become apparent. These difficulties can be overcome by strict adherence to proper scanning techniques, which involve transducer selection, tissue attenuation compensation, and alterations in patient position. The availability of variable-dynamic-range signal processing and the use of real-time scanning can further increase one's confidence in the correct interpretation of these masses. The primary criteria for determining that a mass is fluid-filled have been expanded to include the presence of reverberation echoes, the "lateral shades" sign, and the presence of septations. In the past, hypoechoic masses with low-level internal echoes were termed "complex". Both fluid-filled masses and solid masses may fall into this category. By use of the sonographic criteria, an attempt should be made to determine whether a mass is primarily fluid-filled or solid. Specific anatomic locations and pathologic conditions in which differential diagnosis may be difficult are illustrated; these include abdominal masses, hepatic and renal masses, and pelvic masses.

Abdominal Neoplasms↗
PubMed1977

The frequency of P1 transduction of the genes of Escherichia coli as a function of chromosomal position: preferential transduction of the origin of replication.

The frequencies with which the generalized transducing phage P1 transduced 26 selected markers on the E. coli. chromosome were measured. The frequencies were found to vary relative to argH+ = 1 from a maximum of 6.8 near the origin of replication to a minimum of 0.23 for a marker not far from the terminus. The low frequencies obtained for some markers were shown not to result from poor expression under the selective conditions employed. When plotted as a function of marker position on the chromosome the frequencies were found to exhibit a series of peaks and troughs which correspond to those in gene density noted by Bachmann et al. (1976). The possible relationship of these results to the structure of the E. coli chromosome and to the mechanism of generalized transduction are discussed.

Chromosome Mapping↗
PubMed2027

Lentiviral Transduction of Embryonic Stem Cells.

Lentiviral vectors provide an efficient and reliable method for stable gene knockdown in embryonic stem cells (ESCs) through RNA interference. Here, we describe a detailed protocol for lentiviral transduction of mouse ESCs using lentiviral shRNA expression vectors. The protocol encompasses lentiviral particle production in HEK-293T packaging cells, determination of viral titer, transduction of ESCs cultured under feeder-free conditions, and selection of stably transduced cells. Additionally, we describe methods for evaluating transduction efficiency using fluorescence microscopy and flow cytometry, as well as for assessing gene knockdown efficacy by quantitative real-time PCR (Q-RT-PCR). This protocol is suitable for functional genomic studies in pluripotent stem cells and can be adapted for other difficult-to-transfect cell types.

Lentivirus↗
PubMed1979

Isolation and characterization of transducing phage coding for sigma subunit of Escherichia coli RNA polymerase.

A transducing phage has been isolated with codes for the sigma subunit of Escherichia coli RNA polymerase. Transducing phage were selected from E. coli shotgun collections of HindIII or Sac I fragments cloned into Charon 25, a new bacteriophage lambda vector that is capble of forming lyosogens at high temperature. Transduction of an E. coli strain carrying a temperature-sensitive mutation in the sigma gene was used for the selection. The positions of restriction sites for Sac I, HindIII, Xho I, Bgl II, and Kpn I in the cloned bacterial DNA segments were determined. Phage containing the HindIII fragment complement both primase (dnaG) and sigma (rpoD) whereas those containing the Sac I fragment complement only sigma. Results of analyses of the proteins made both in vivo after infection of UV-irradiated cells and in vitro in a coupled transcription/translation system suggest that a Sac I site separates the promoter for sigma from the sigma structural gene. The direction of transcription of sigma was determined to be clockwise with respect to the E. coli genetic map.

Bacterial Proteins↗
PubMed1977

Escherichia coli K-12 mutants hyperproducing chromosomal beta-lactamase by gene repetitions.

Escherichia coli K-12 ampicillin-resistant mutants hyperproducing chromosomal beta-lactamase arose spontaneously from strains carrying ampA1 ampC(+). Such mutants were found even in a recA background. Two Amp(r)-100 strains were analyzed genetically. The Amp(r)-100 resistance level of both strains could be transduced by direct selection for ampicillin resistance. Several classes of ampicillin-resistant transductants were found that differed from one another in the beta-lactamase activity and the ampicillin resistance mediated by an ampA1 ampC(+)-carrying strain. The data suggested that beta-lactamase hyperproduction was due to repetitions of the chromosomal amp genes. The size of the repeated region was calculated from cotransduction estimates, using the formula of Wu (Genetics 54:405-410, 1966), and was found to be about 1 min in one strain and 1.5 min in the other. Second-step Amp(r)-400 mutants were isolated from an Amp(r)-100 strain. The resistance of these mutants was apparently also due to repetitions, each mediating a resistance to about 10 mug/ml. Mutants of wild-type strains that were moderately resistant to ampicillin also gave rise to intermediate-resistance classes, suggesting repetitions of the wild-type amp alleles. F' factors hyperproducing chromosomal beta-lactamase by gene repetitions were constructed. They mediated levels of ampicillin resistance comparable to that of naturally occurring resistance plasmids. The expression of beta-lactamase hyperproduction was not affected by the presence of ampA and ampC alleles in trans and did not act in trans on the other alleles.

Alleles↗
PubMed1978

Transposition of a DNA sequence determining kanamycin resistance into the single-stranded genome of bacteriophage fd.

Derivatives of bacteriophages fd which transduce kanamycin resistance were selected after growth of the phage in an E. coli strain that carried transpoon 5 (Tn5). Different clones of transducing phage and their DNAs were characterized by gel electrophoresis, electron microscopy, and by their ability to multiply in the absence of helper phage. Integration of the intact transposon into the full size phage genome was correlated with an increase in size of the phage particle from 0.95 mu to 1.7 mu, and with the appearance in the phage DNA of the stem loop structure characteristic for single-stranded Tn5 DNA. In non-defective phages the site of insertion was mapped by heteroduplex analysis within the intergenic region of the phage genome. Defective transducing phages were characterized as an insertion of Tn5 into a phage gene, and/or as a partial deletion or duplication of phage and transposon DNA. The size of the transducing phage from different defective clones varied from 0.6 mu to 3.0 mu and was directly proportional to the DNA content. These results demonstrate that filamentous bacteriophage are highly capable to replicate and package very different amounts of foreign DNA.

Coliphages↗
PubMed1977

Effects of auditory masking on lip movements during speech.

This study describes the effects of unpredictably presented masking noise on selected parameters of lip movement during speech. Four normal subjects produced a simple speech utterance while vertical displacements of the upper and lower lips were transduced and recorded. During selected productions a masking noise was presented dichotically at one of two distinct phases of lip movement. Analysis of the resulting data revealed the following for all subjects: (1) significant changes in the duration, displacement, and velocity of lip movement caused by masking and (2) significant reductions in the velocity of lip movement occurring approximately 50 msec after the onset of the masker. Possible explanations of these and certain speaker-dependent effects are discussed.

Acoustic Stimulation↗
PubMed1978

Identification of a host protein necessary for bacteriophage morphogenesis (the groE gene product).

Mutations in the groE gene of Escherichia coli, which block the correct assembly of the phage lambda head, have been previously described. Many groE mutations exert pleiotropic effects, such as inability to propagate phages T4 and T5 and inability to form colonies at 43 degrees. With the help of the EcoRI and HindIII restrictionenzymes and the appropriate phage vectors, we have constructed two lambda transducing phages, called W3 and H18, that carry the groE+ bacterial gene. Upon lysogenization by phage H18 the groE bacterial mutants recover their gro+ phenotype for both phage growth and the ability to form colonies at 43 degrees. We have identified the groE+ bacterial gene product as a protein of 65,000 molecular weight. Mutants of the W3 transducing phage that were selected on the basis of their ability to propagate on some groE mutant hosts induce the synthesis of a groE protein with altered electrophoretic mobility.

Bacterial Proteins↗
PubMed1978

The effects of Triton-detergents on the stretch receptor of the crayfish.

The effects of the non-ionic detergents Triton X-45 and Triton X-100 on the action potential and the receptor potential of the stretch receptor neuron of the crayfich Astacus fluviatilis was studied with intracellular recording technique. Membrane currents were measured with voltage clamp technique. Both detergents blocked the action potential in 20--30 min at concentrations of 60--80 microgram. Following blocking of spike electrogenesis the receptor potential evoked by strech was obtained in isolation. With prolonged exposure of the neuron to the detergents there was a slowly developing reduction of the receptor potential and after 60--90 min no response to stretch could be obtained. These effects were produced without any significant change of the resting membrane potential. Following return to normal saline the responsiveness to stretch was completely restored in 80--100 min. Measurements with voltage clamp techinque showed that the passive membrane properties were little affected by the two detergents. The stretch induced current on the other hand was severely depressed and almost abolished with prolonged exposure. The experimental results suggest that non-ionic detergents block the spike electrogenesis and the transducer action by a selective action on the sodium channels of the membrane of the receptor neuron.

Action Potentials↗
PubMed1979

Acoustical analysis of the auditory system of the cricket Teleogryllus commodus (Walker).

The basic auditory physiology of crickets, and particularly of Teleogryllus commodus (Walker) is examined and its behavior simulated by electrical analog networks, beginning from the simplest possible model and progressing by stages to the full system found in the real insect. It is found that the attenuation of sound in the auditory trachea plays a crucial role in the mechanism for directional hearing in even the simplest model and that the tracheal diameter is in fact appropriate to produce the desired attenuation. In a more complex model in which it is recognized that the auditory system probably responds to pressure changes in the tracheal sacs underlying the tympana rather than simply to tympanic motion, it is found that the phase shift produced by the combined effects of the central septum and the adjoining cavities leading to the spiracles is also important to hearing directionality. The final model which includes both tympana and spiracles is able to simulate both the hearing directionality and, in part, the frequency selectivity of the system. It appears, however, that a large measure of the observed frequency selectivity is due to some form of selectivity in the neural transducers themselves rather than in the simple acoustic components of the system.

Acoustics↗
PubMed1977

Specialized transducing phage for the initiation factor 3 gene in Escherichia coli.

A previously isolated thermosensitive mutant [Springer, M., Graffe, M. & Grunberg-Manago, M. (6977) Mol. Gen. Genet. 151, 17-26] exhibits two defects in vitro, one in the initiation factor IF3 and the other in the L-phenylalanine: tRNA-Phe ligase (EC 6.1.1.20). Specialized lambda transducing phages that transduced this mutant to thermoresistance were selected. In vitro studies showed that the transductants had a normal IF3 activity. One of these transducing phages was shown to code for a protein synthesized under the control of Escherichia coli promoters, which has the same molecular weight as IF3. This protein crossreacts specifically with IF3 antisera and comigrates with pure IF3 in a two-dimensional gel system.

Bacterial Proteins↗
PubMed1978

Nonsense and insertion mutants in the relA gene of E. coli: cloning relA.

We have made use of lysogens of a specialized transducing bacteriophage, lambdapyrG+ relA+, to select nonsense (relAnon) and insertion (relAins) mutations in the relA gene. Three independent relAnon mutants were isolated on the phage. In all three, the relaxed phenotype was suppressed by supD, supE, supF or sup6. Three independent relAins mutants were isolated, all containing an insertion element (probably IS2) in an apparently identical location in the relA gene. Polyacrylamide gel electrophoretic analysis of peptides synthesized by the phages in ultraviolet lightkilled host cells revealed that no stringent factor was coded for by either the relAins or relAnon phages (the latter in a sup+ cell); stringent factor was detected when the relAnon phages were used in a similar experiment with supD or supE host cells. The relAnon and relAins mutations could be crossed in haploid form in the E. coli chromosome. These recombinants grew with a normal doubling time, had a ppGpp pool which was between 70 and 100% compared with the classical relA strain, and underwent a normal carbon source shift-down. A restriction endonuclease map of the pyrG relA region of the specialized transducing phage is presented in which the position of the insertion element (recognized by a novel Hind III-cut site) defines the position of the relA gene. This position was verified by an analysis of the structure of five plasmids formed by cloning portions of the region in the pBR322 cloning vehicle. Our results indicate that the relA gene is not an essential cellular function, that there might be a second mechanism for the synthesis of basal level ppGpp in the cell and that the sole function of the relA gene is apparently the high level ppGpp synthesis triggered in response to deacylated tRNA.

Bacterial Proteins↗
PubMed1976

Genetic mapping of xthA, the structural gene for exonuclease III in Escherichia coli K-12.

The genes xthA, pncA, and pabB were ordered relative to others by two- and three-factor transductional crosses with bacteriophage P1. The genes studied span 2 min (2%) of the genetic map of Escherichia coli K-12 in the clockwise sequence pheS-pfkB-xthA-pncA-gap-pabB-fadD. Eleven independently derived xth mutations were examined; all were known to affect exonuclease III and its associated endonuclease II activity, and all were mapped in the xthA region. pncA mutations were found to confer resistance to 6-aminonicotinamide, whereas some pheS mutations are known to specify resistance to p-fluorphenylalanine. xth mutations were readily transferred into other strains by selecting for these co-transducible drug resistance markers.

6-Aminonicotinamide↗
PubMed2025

Exploring phage-host interactions in Burkholderia cepacia complex bacterium to reveal host factors and phage resistance genes using CRISPRi functional genomics and transcriptomics.

Complex interactions of bacteriophages with their bacterial hosts determine phage host range and infectivity. While phage defense systems and host factors have been identified in model bacteria, they remain challenging to predict in non-model bacteria. In this paper, we integrate functional genomics and transcriptomics to investigate phage-host interactions, revealing active phage resistance and host factor genes in Burkholderia cenocepacia K56-2. Burkholderia cepacia complex species are commonly found in soil and are opportunistic pathogens in immunocompromised patients. We studied infection of B. cenocepacia K56-2 with Bcep176, a temperate phage isolated from Burkholderia multivorans. A genome-wide dCas9 knockdown library targeting B. cenocepacia K56-2 was constructed, and a pooled infection experiment identified 63 novel genes or operons coding for candidate host factors or phage resistance genes. The activities of a subset of candidate host factor and resistance genes were validated via single-gene knockdowns. Transcriptomics of B. cenocepacia K56-2 during Bcep176 infection revealed that expression of genes coding for host factor and resistance candidates identified in this screen was significantly altered during infection by 4 h post-infection. Identifying which bacterial genes are involved in phage infection is important to understand the ecological niches of B. cenocepacia and its phages, and for designing phage therapies.IMPORTANCEBurkholderia cepacia complex bacteria are opportunistic pathogens inherently resistant to antibiotics, and phage therapy is a promising alternative treatment for chronically infected patients. Burkholderia bacteria are also ubiquitous in soil microbiomes. To develop improved phage therapies for pathogenic Burkholderia bacteria, or engineer phages for applications, such as microbiome editing, it's essential to know the bacterial host factors required by the phage to kill bacteria, as well as how the bacteria prevent phage infection. This work identified 65 genes involved in phage-host interactions in Burkholderia cenocepacia K56-2 and tracked their expression during infection. These findings establish a knowledge base to select and engineer phages infecting or transducing Burkholderia bacteria.

Bacteriophages↗