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Y Komeda

Publications and source records attributed to Y Komeda.

At least 73 records · Page 4Linked to original sources

Transcriptional control of flagellar genes in Escherichia coli K-12.

Autoregulation of the expression of flagellar genes was investigated by the technique of operon fusion. The results suggested that the flaU gene is a repressor and the flaD gene is an activator of transcription of the hag, flaS, and Mocha operons. The action of the putative flaU repressor appears to be masked by its interaction with other flagellar proteins during assembly; thus, repression is apparent only when the interacting proteins are absent. This hypothesis is supported by the phenotype of an unusual flaU mutant, which represses even though it is unable to promote flagellar assembly. Presumably, the mutant synthesizes a repressor whose activity is no longer masked by interaction with other flagellar proteins.

Escherichia coli↗

[A case of acute epididymitis in infancy].

We report a 3-month-old boy who presented with acute right scrotal swelling that was diagnosed to have epididymitis at surgical exploration. Urine culture and urinalysis were negative and IVP revealed no abnormality. White blood count and symptom were improved with chemotherapy. Acute epididymitis in infancy is very rare and this case seems to be the 16th case in Japan.

Acute Disease↗

[Six cases of spontaneous peripelvic extravasation from the renal pelvis].

The diagnosis and management of 6 patients with spontaneous non-traumatic urinary extravasation are reported. It is important to distinguish extravasation of the fornical backflow type from that owing to frank rupture of the diseased renal pelvis. In Japan, 26 cases of spontaneous peripelvic extravasation and 29 cases of spontaneous pelvic rupture have been reported. Most of them were caused by calculous ureteral obstruction. Many of the cases of spontaneous peripelvic extravasation could be managed conservatively, and many of the cases of spontaneous pelvic rupture often required surgical treatment due to the patient's clinical condition, the persistence of obstruction or extravasation, or the presence of complication of extravasation such as urinoma or abscess. All of our 6 cases were managed conservatively and had no complications.

Adult↗

Synthesis of flagellin and hook subunit protein in flagellar mutants of Escherichia coli K12.

We have examined Escherichia coli K12 flagellar mutants affected in each of 29 different loci for the synthesis of flagellin and hook subunit protein. Immune precipitation experiments were employed by treating cell extracts with antiserum against each protein. Flagellin was synthesized in mutants defective in genes flaS , flaT, flaU and flbC . The flaE and flaZ mutants produced small amounts of flagellin, while all the other mutants failed to produce any detectable amount of flagellin. Hook subunit protein was found in most mutants including those defective in genes flaA , flaB , flaC, flaD , flaE , flaG , flaH , flaL , flaM , flaN , flaO , flaP, flaQ , flaR , flaS , flaT, flaU , flaV , flaW , flaX, flaY , flaZ , flbA , flbC , flbD , and hag but not in mutants of flaK , flaI , and flbB . The results conform to the predictions made by our previous indirect gene fusion study ( Komeda 1982).

Bacterial Proteins↗

Bacteriophage chi sensitivity and motility of Escherichia coli K-12 and Salmonella typhimurium Fla- mutants possessing the hook structure.

The production of hook protein and flagellin in 29 Fla- mutants of Escherichia coli K-12 was determined by the complement fixation assay. Six mutants produced hook protein, and four of them also produced flagellin. A flaE mutation was introduced into these fla mutants carrying the hook structure. All of these mutants made polyhooks and were used as hosts for a newly isolated host-range mutant of chi phage that has a high affinity for the hook structure. All except one mutant produced significant amounts of progeny phages. A flaD flaE double mutant was that exception which did not yield significant amounts of progeny by the phage propagation method. All of the flaE double mutants produced comparable amounts of polyhooks, and no qualitative difference was detected between chi-sensitive and chi-insensitive mutants by the complement fixation assay. Accordingly, it was thought that the polyhook of the flaD flaE mutant had a mechanical defect for chi phage infection. This assumption was confirmed by tethered-cell experiments; the flaD flaE mutant did not rotate. These results are well explained by a proposed regulation pathway of flagellar genes. flaE mutants can express other genes which govern the final step of the flagellar morphogenesis, whereas flaD mutants cannot rotate, possibly because the mocha operon is not expressed. The results obtained in E. coli were also found to be applicable to Salmonella typhimurium.

Bacterial Proteins↗

[Study of bulbocavernosus reflex using electrostimulation electromyography].

Evoked response of bulbocavernosus reflex was taken in 8 normal control individuals and in 15 abnormal voiding patients (8 benign prostatic hyperplasia, 4 diabetic neuropathy, 3 supranuclear neurogenic bladder). the latency in control group showed 31.5 +/- 5.0 msec duration after penile stimulation and 65.4 +/- 11.0 msec duration after posterior urethral stimulation. The latency in diabetic neuropathy group was obviously prolonged but not that in the supranuclear neurogenic bladder group. This modern neurophysiological electrostimulation method provides simple and accurate information of bulbocavernosus reflex.

Adult↗

[The dynamics of corticosteroids levels in maternal and fetal plasma].

In order to elucidate the clinical implications of cortisol levels in pregnancy, maternal and fetal serum levels of cortisol during pregnancy were determined. The maternal serum level of cortisol gradually increased with advancing gestational age, and it reached a 3 to 4 times higher level in the late stage of pregnancy than that in non-pregnant state. The maternal serum level of cortisol during delivery increased with the length of the period of labor, and immediately after the delivery it reached the peak value of 60-80 micrograms/dl. According to the mode of delivery, the cortisol level increased with the accumulation of stress on the maternal side. The value was highest in the case of vacuum extraction and lowest in elective cesarean section. The cortisol level in umbilical arterial blood in normal delivery was not significantly elevated in comparison with that in vacuum extraction, oxytocin induced delivery and elective cesarean section. The cortisol level in umbilical blood of anencephaly did not differ from that of normal neonate, indicating that cortisol in umbilical blood may originate mainly on the maternal side. In the postpartum period, the serum cortisol concentration declined to the level observed in the late stage of pregnancy after 2 days, and further decreased to the level in the non-pregnant state after 6 days. The present studies suggest that the increase in the serum concentration of cortisol at delivery is a result of maternal stresses in labor, and that direct involvement of cortisol in the controlling mechanism of puerperium is unlikely. The increased function of the adrenal cortex in the late stage of pregnancy and at delivery may stimulate fetal pulmonary maturity and provide maternal and fetal adaptability to stresses in intrapartum bleeding and shock during labor. It is essential to understand pituitary-adrenal cortex functions in prenatal and fetal care in pregnancy.

Anencephaly↗

[Studies on the secretion of human placental lactogen from human trophoblastic tissues (author's transl)].

In order to study the secretory mechanism of human placental lactogen (hPL:hCS) from trophoblastic tissues, tissue culture and new placental perifusion systems were developed and used to clarify the effect of various substances on the secretion of hPL. These substances were (1), metal ions([Ca2+], [K+], [mg2+], [Na+]); (2) growth hormone and prolactin releasing or inhibiting factors (arginine, TRH, somatostatin, dopamine); (3) LH-RH, dibutyryl cyclic-AMP which stimulates hCG secretion; (4) prostaglandins F2 alpha and E2, bradykinin; (5) EGF and insulin which have the receptors in the placenta; (6) glucose. It was found that most of the substances examined had no effect on the secretion of hPL, except dopamine and glucose. The effect of dopamine in the tissue culture system is dose-dependent. At high concentrations dopamine slightly inhibited hPL secretion(5mM: 38.6 +/- 15.0 and 10mM; 35.7 +/- 19.0 micrograms/g wet tissue) compared with the control (63.2 +/- 29.8 microgram/g wet tissue). However, these effects may be due to the deviation of pH in the medium from the direct addition of dopamine hydrochloride. At a low concentration(1mM) it was observed to have a rather stimulatory effect (125.7 +/- 18.0 micrograms/g wet tissue, p less than 0.05), but in the perifusion system, this effect could not be observed. The addition of glucose in the perifusion system gave a slightly higher hPL secretion than that of the control. Perhaps this resulted from increased cell activity rather than a stimulatory effect. an incorporation experiment of [3H] leucine was also carried out to study the biosynthesis of hPL. Newly synthesized ([3H]-labelled) hPL was secreted into the medium within two hours. Furthermore, a labelled larger molecular weight substance together with the tritiated hPL was also observed on a Sephadex G-100 gel chromatography. These labelled substances were immunoprecipitable using an anti-hPL serum, indicating that the substances contain the same immunological determinants. This result indicates that the larger molecular substance may represent the biosynthetic precursor or the aggregate of hPL. These data indicate that the secretion of hPL has a unique mechanism, different from GH and PRL, and may be self regulated.

Arginine↗

Fusions of flagellar operons to lactose genes on a mu lac bacteriophage.

Previous studies have defined 29 genes necessary for synthesis of the Escherichia coli flagellar apparatus. This study analyzed the transcriptional control of flagellar genes, using Mu d (Apr lac) phage to generate flagellar mutants by insertion. These mutants contained operon fusions of flagellar genes to the lac genes of the Mu d phage and allowed the measurement of flagellar operon expression by detection of beta-galactosidase activity. These fusion mutants expressed the enzyme activity constitutively, and an autogenous regulation mechanism was not revealed. Lambda transducing phages carrying these chromosomal fla-lac fusions were also isolated and used to examine the effect of different fla mutations on expression of each flagellar operon. The results showed that flagellar operons are divided into six classes; (class 1) the flbB operon, which controls all of the other flagellar operons; (class 2) the flaU and flbC operons, which are controlled by the flbB operon gene products and are not required for the expression of other Fla operons; (class 3) the flbA, flaG, flaD, flaN, flaB, and flaA operons, which are under flbB operon control and are required for the expression of other fla operons; (class4) the flaZ operon, which is controlled by the gene products of the group 1 and 3 operons and is required for hag transcription; (class 5) the mocha and flaS operons, which are controlled by the gene products of the group 1 and 3 operons; and (class 6) the hag operon. These results are discussed with respect to the possible assembly sequence of the fla gene products.

Coliphages↗

Incomplete flagellar structures in Escherichia coli mutants.

Escherichia coli mutants with defects in 29 flagellar genes identified so far were examined by electron microscopy for possession of incomplete flagellar structures in membrane-associated fractions. The results are discussed in consideration of the known transcriptional interaction of flagellar genes. Hook-basal body structures were detected in flaD, flaS, flaT, flbC, and hag mutants. The flaE mutant had a polyhook-basal body structure. An intact basal body appeared in flaK mutants. Putative precursors of the basal body were detected in mutants with defects in flaM, flaU, flaV, and flaY. No structures homologous to the flagellar basal body or its parts were detected in mutants with defects in flaA, flaB, flaC, flaG, flaH, flaI, flaL, flaN, flaO, flaP, flaQ, flaR, flaW, flaX, flbA, flbB, and flbD. One flaZ mutant had an incomplete flagellar basal body structure and another formed no significant structure, suggesting that flaZ is responsible for both basal body assembly and the transcription of the hag gene.

Bacterial Proteins↗

Definition of additional flagellar genes in Escherichia coli K12.

Twenty-nine flagellar genes in Escherichia coli K12 have previously been assigned to three regions of the genome. Flagellar region I is located between pyrC and ptsG, region II between aroD and uvrC, and region III between uvrC and his. In this study, flagellar mutants in Escherichia coli K12 were obtained by selection for resistance to the flagellotropic phage, chi. They were analyzed in complementation tests using P1 phage-mediated transduction. In addition to the fla genes already described, eight more flagellar genes were identified. This analysis defined six more fla genes in region I (flaU, etc.), one more in region II (flbB) and one more in region III (flbC). Region I was shown to include at least 12 fla cistrons. Complementation analysis with polar Mu phage-induced Fla- mutants and with lambda fla phage defined four transcriptional units in region I. These were: flaU, flbA-flaW-flaV-flaK-flaX-flaL-flaY-flaM, flaZ and flaS- flaT, with transcription proceeding from left to right. The flB gene was found to be part of an operon: flB-flaI in region II. In region III, a previously unidentified gene flbC was located between hag and flaN.

Chromosome Mapping↗

Regulation of expression of the flagellin gene (hag) in Escherichia coli K-12: analysis of hag-lac gene fusions.

Previous studies have defined 28 genes necessary for the synthesis of the flagellar apparatus of Escherichia coli K-12. This study analyzed the influence of the flagellar genes on the expression of the hag gene (structural gene for flagellin). To this end, a hag::Mu d(Apr lac) mutant which had the lac genes fused to the promoter of the hag gene was constructed. This allowed the measurement of hag gene expression by detection of beta-galactosidase activity. The following observations were made. (i) The hag gene was expressed constitutively in Fla+ cells. (ii) hag gene expression was positively regulated by flaA, FLAB, flaC, flaD, flaE, flaG, flaH, flaI, flaK, flaL, flaM, flaN, flaO, flaP, flaQ, flaR, flaV, flaW, flaX, flaY, flaZ, flbA, and flbB genes.hag-lac expression was not observed in strains with these fla mutations. (iii) The hag gene was expressed in mutants with flaS, flaT, flaU, and flbC defects. Therefore, these genes were not involved in regulation of hag gene transcription.

Bacterial Proteins↗

Identification of the structural gene for the hook subunit protein of Escherichia coli flagella.

Previous studies showed that the structural gene for the flagellar hook subunit protein (molecular weight 42,000) was one of a group of flagellar genes located on the Escherichia coli genome near pyrC. Several lines of evidence indicate that the flaK gene is the structural gene for the hook subunit protein. Fla+ strains that were insensitive to chi infection could be isolated as revertants of an FlaK- amber mutant strain but from no other Fla- strain. The hook subunit proteins isolated from such chi-sensitive revertants of the FlaK- strain were shown to be antigenically and electrophoretically different from the hook protein isolated from the wild-type strain. Thus, reversion of a mutation in the flaK gene resulted in alteration of the structure of the hook protein. Furthermore, in programming experiments with hybrid lambda containing flagellar genes, lambdafla with flaK genetic activity programmed the synthesis of a 42,000-molecular weight protein, whereas lambdafla without flaK genetic activity did not.

Bacterial Proteins↗

Genes for the hook-basal body proteins of the flagellar apparatus in Escherichia coli.

Of the more than 30 genes required for flagellar function, 6 are located between pyrC and ptsG on the Escherichia coli genetic man. This cluster of genes is called flagellar region I. Four-point transductional crosses were used to establish the position and order of the region I flagellar genes with respect to the outside markers ptsG and pyrC. Bacteriophage lambda-E. coli hybrids that contained most of the genes necessary for flagellar formation were constructed. The properties of specific hybrids that carried the region I fla genes were examined by genetic complementation and by measuring the capacity of the hybrids to direct the synthesis of specific polypeptides. The results of these tests with lambda hybrids and with a series of deletion mutations derived from the lambda hybrids demonstrated the existence of at least six flagellar-specific cistrons. These directed the synthesis of polypeptides with the following apparent molecular weights: flaV, 11,000; flaK, 42,000; flaL, 30,000 and 27,000; flaM, 38,000; flS, 60,000; and flaT, 35,000. Plasmid ColE1-E. coli hybrids with region I flagellar genes were also used to program the synthesis of polypeptides in minicell-producing strains. The polypeptides synthesized in these experiments were identical to polypeptides of the hook-basal body structure and helped to confirm the assignment of genes to specific polypeptides. The synthesis of all of these polypeptides was regulated by the same mechanism that regulates the synthesis of other flagellar-related structural components.

Bacterial Proteins↗

Effects of galU mutation on flagellar formation in Escherichia coli.

Two mutants of Escherichia coli strictly deficient in uridine-diphosphoglucose pyrophosphorylase activity (galU) were found to have very small numbers of flagellar filaments and hooks. In these mutants, both the rate of flagellin (flagellar protein) synthesis and the amount of messenger ribonucleic acid specific for flagellin were considerably lower than in the parental strains. Motile revertants from the galU mutants were isolated and were found to carry a suppressor mutation, which was mapped in the flaH cistron. These strains formed swarms under conditions of catabolite repression; their intracellular concentration of cyclic adenosine 5'-monophosphate was the same as that in the parental strains. These results suggest that the outer membrane affects flagellar formation through the flaH gene product.

Escherichia coli↗