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Mutants Deficient (mutant + deficient)
Selected AbstractsActivity of Serratia plymuthica IC1270 gene chiA promoter region in Escherichia coli mutants deficient in global regulators of transcriptionJOURNAL OF BASIC MICROBIOLOGY, Issue 6 2005I. A. Khmel To study the regulation of expression of the Serratia plymuthica gene chiA encoding a 58-kDa endochitinase, its 586-bp-long upstream regulatory region was cloned, sequenced and fused to a promoterless lac operon in phage ,RS45 to obtain a single-copy transcriptional fusion (PF1chiA - lac ) in lysogens of Escherichia coli wild-type strains or their mutants deficient in various global regulators of transcription. The level of PF1chiA - lac expression increased about 20- and 90-fold, respectively, in E. coli K12 ,hns and double ,hns stpA mutants deficient in H-NS, and in both H-NS and StpA DNA-binding histone-like proteins, as compared to levels in the wild-type strain. In a ,lrp mutant deficient in the leucine-responsive transcriptional regulator Lrp, the level of PF1chiA - lac expression increased only up to threefold, whereas even smaller differences relative to the wild-type strain were observed in rpoS and ,crp mutants deficient in the ,S subunit of RNA polymerase and catabolite-repression protein (CRP), respectively. Deletion of the inverted-repeat sequences and curved DNA regions located in the upstream region of chiA essentially did not influence strain IC1270's chiA promoter activity in E. coli . (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Isolation and characterization of a transposon mutant of Pseudomonas aeruginosa affecting uptake of dibenzothiophene in n -tetradecaneLETTERS IN APPLIED MICROBIOLOGY, Issue 2 2003K. Noda Abstract Aims: Isolation and characterization of a transposon mutant of Pseudomonas aeruginosa affecting the uptake of dibenzothiophene (DBT) in n -tetradecane (n -TD). Methods and Results: The dsz desulphurization gene cluster from Rhodococcus erythropolis KA2-5-1 was transferred to the chromosome of P. aeruginosa NCIMB9571 using a transposon vector. A recombinant (named PARM1) was obtained which was able to desulphurize DBT in water, but not in n -TD. Conclusions: PARM1 is a mutant deficient in a DBT transport system operational in n -TD. This transport system is independent of rhamnolipids and of the n -alkane transport system. Significance and Impact of the Study:Pseudomonas aeruginosa NCIMB9571 seems to have a specific system of transporting hydrophobic compounds such as DBT in oil. [source] CO2 limitation induces specific redox-dependent protein phosphorylation in Chlamydomonas reinhardtiiPROTEINS: STRUCTURE, FUNCTION AND BIOINFORMATICS, Issue 9 2006Maria V. Turkina Abstract Acclimation of the green alga Chlamydomonas reinhardtii to limiting environmental CO2 induced specific protein phosphorylation at the surface of photosynthetic thylakoid membranes. Four phosphopeptides were identified and sequenced by nanospray quadrupole TOF MS from the cells acclimating to limiting CO2. One phosphopeptide originated from a protein that has not been annotated. We found that this unknown expressed protein (UEP) was encoded in the genome of C.,reinhardtii. Three other phosphorylated peptides belonged to Lci5 protein encoded by the low-CO2 -inducible gene,5 (lci5). The phosphorylation sites were mapped in the tandem repeats of Lci5 ensuring phosphorylation of four serine and three threonine residues in the protein. Immunoblotting with Lci5-specific antibodies revealed that Lci5 was localized in chloroplast and confined to the stromal side of the thylakoid membranes. Phosphorylation of Lci5 and UEP occurred strictly at limiting CO2; it required reduction of electron carriers in the thylakoid membrane, but was not induced by light. Both proteins were phosphorylated in the low-CO2 -exposed algal mutant deficient in the light-activated protein kinase,Stt7. Phosphorylation of previously unknown basic proteins UEP and Lci5 by specific redox-dependent protein kinase(s) in the photosynthetic membranes reveals the early response of green algae to limitation in the environmental inorganic carbon. [source] Folic acid utilisation related to sulfa drug resistance in Saccharomyces cerevisiaeFEMS MICROBIOLOGY LETTERS, Issue 2 2001Ann M. Bayly Abstract Saccharomyces cerevisiae mutants deficient in folate synthesis have been constructed and employed to study the utilisation of exogenous folates in yeast. One mutant specifically lacked dihydropteroate synthase while the second lacked dihydrofolate synthase. Exogenous folinic acid restored optimal growth to both strains. Folic acid did not generally rescue growth but spontaneous isolates capable of utilising folic acid were selected. The folic acid synthesis pathway in the folate utilising isolates was restored via transformation with FOL1 or FOL3 expression plasmids and transformants were tested for resistance to sulfamethoxazole (SMX). The presence of elevated levels of folic acid led to greatly reduced SMX sensitivity regardless of whether strains were folate utilisers or not. [source] Involvement of gp130-associated cytokine signaling in Müller cell activation following optic nerve lesionGLIA, Issue 7 2010Matthias Kirsch Abstract Ciliary neurotrophic factor (CNTF) and the related cytokine leukemia inhibitory factor (LIF) have been implicated in regulating astrogliosis following CNS lesions. Application of the factors activates astrocytes in vivo and in vitro, and their expression as well as their receptors is upregulated after brain injury. Here, we investigated their function by studying Müller cell activation induced by optic nerve crush in CNTF- and LIF-deficient mice, and in animals with deficiencies in cytokine signaling pathways. In the retina of CNTF,/, mice, basal GFAP expression was reduced, but unexpectedly, injury-induced upregulation in activated Müller cells was increased during the first 3 days after lesion as compared to wild-type animals and this corresponded with higher phosphorylation level of STAT3, an indicator of cytokine signaling. The observation that LIF expression was strongly upregulated in CNTF,/, mice but not in wild-type animals following optic nerve lesion provided a possible explanation. In fact, additional ablation of the LIF gene in CNTF/LIF double knockout mice almost completely abolished early lesion-induced GFAP upregulation in Müller cells and STAT3 phosphorylation. Early Müller cell activation was also eliminated in LIF,/, mice, despite normal CNTF levels, as well as in mutants deficient in gp130/JAK/STAT signaling and in conditional STAT3 knockout mice. Our results demonstrate that LIF signaling via the gp130/JAK/STAT3 pathway is required for the initiation of the astrogliosis-like reaction of retinal Müller cells after optic nerve injury. A potential role of CNTF was possibly masked by a compensatory increase in LIF signaling in the absence of CNTF. © 2010 Wiley-Liss, Inc. [source] Activity of Serratia plymuthica IC1270 gene chiA promoter region in Escherichia coli mutants deficient in global regulators of transcriptionJOURNAL OF BASIC MICROBIOLOGY, Issue 6 2005I. A. Khmel To study the regulation of expression of the Serratia plymuthica gene chiA encoding a 58-kDa endochitinase, its 586-bp-long upstream regulatory region was cloned, sequenced and fused to a promoterless lac operon in phage ,RS45 to obtain a single-copy transcriptional fusion (PF1chiA - lac ) in lysogens of Escherichia coli wild-type strains or their mutants deficient in various global regulators of transcription. The level of PF1chiA - lac expression increased about 20- and 90-fold, respectively, in E. coli K12 ,hns and double ,hns stpA mutants deficient in H-NS, and in both H-NS and StpA DNA-binding histone-like proteins, as compared to levels in the wild-type strain. In a ,lrp mutant deficient in the leucine-responsive transcriptional regulator Lrp, the level of PF1chiA - lac expression increased only up to threefold, whereas even smaller differences relative to the wild-type strain were observed in rpoS and ,crp mutants deficient in the ,S subunit of RNA polymerase and catabolite-repression protein (CRP), respectively. Deletion of the inverted-repeat sequences and curved DNA regions located in the upstream region of chiA essentially did not influence strain IC1270's chiA promoter activity in E. coli . (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source] Iron uptake is essential for Escherichia coli survival in drinking waterLETTERS IN APPLIED MICROBIOLOGY, Issue 1 2006D. Grandjean Abstract Aims:, The aim of this study was to elucidate if the need for iron for Escherichia coli to remain cultivable in a poorly nutritive medium such as the drinking water uses the iron transport system via the siderophores. Methods and Results:, Environmental strains of E. coli (isolated from a drinking water network), referenced strains of E. coli and mutants deficient in TonB, an essential protein for iron(III) acquisition, were incubated for 3 weeks at 25°C, in sterile drinking water with and without lepidocrocite (, -FeOOH), an insoluble iron corrosion product. Only cells with a functional iron transport system were able to survive throughout the weeks. Conclusions:, The iron transport system via protein TonB plays an essential role on the survival of E. coli in a weakly nutritive medium like drinking water. Significance and Impacts of the Study:, Iron is a key parameter involved in coliform persistence in drinking water distribution systems. [source] Thymineless death is associated with loss of essential genetic information from the replication originMOLECULAR MICROBIOLOGY, Issue 6 2010Dipen P. Sangurdekar Summary Thymine starvation results in a terminal cellular condition known as thymineless death (TLD), which is the basis of action for several common antibiotics and anticancer drugs. We characterized the onset and progression of TLD in Escherichia coli and found that DNA damage is the only salient property that distinguishes cells irreversibly senesced under thymine starvation from cells reversibly arrested by the nucleotide limitation. The damage is manifested as the relative loss of genetic material spreading outward from the replication origin: the extent of TLD correlates with the progression of damage. The reduced lethality in mutants deficient in the RecFOR/JQ repair pathway also correlates with the extent of damage, which explains most of the observed variance in cell killing. We propose that such spatially localized and persistent DNA damage is the consequence of transcription-dependent initiation of replication in the thymine-starved cells and may be the underlying cause of TLD. [source] Why does elevated CO2 affect time of flowering?NEW PHYTOLOGIST, Issue 2 2009An exploratory study using the photoperiodic flowering mutants of Arabidopsis thaliana Summary ,,Evidence is accumulating that the effect of CO2 on time of flowering involves interactions with photoperiod, but the basis for this interaction is unclear. Here, which components of the photoperiod flowering pathway account for this interaction in Arabidopsis thaliana were examined. ,,Ten mutants deficient in particular loci in the photoperiod pathway, as well as the wild type, were grown under short and long days at either ambient or elevated CO2. Leaf number at flowering and the number of days required for induction of flowering were determined. ,,Elevated CO2 interacted with both the photoreceptors and the subsequent transduction reactions in the photoperiod pathway. The direction and magnitude of the effects varied with photoperiod. Elevated CO2 also affected flowering by increasing rate of leaf production. ,,The net effect of elevated CO2 on time of flowering varies because CO2 has a complex array of effects on different elements of the developmental pathway leading to flower induction that may either hasten or delay flowering depending upon the influence of other environmental factors such as photoperiod. [source] Reactive oxygen signaling and abiotic stressPHYSIOLOGIA PLANTARUM, Issue 3 2008Gad Miller Reactive oxygen species (ROS) play a dual role in plant biology acting on the one hand as important signal transduction molecules and on the other as toxic by-products of aerobic metabolism that accumulate in cells during different stress conditions. Because of their toxicity as well as their important signaling role, the level of ROS in cells is tightly controlled by a vast network of genes termed the ,ROS gene network'. Using mutants deficient in key ROS-scavenging enzymes, we have defined a signaling pathway that is activated in cells in response to ROS accumulation. Interestingly, many of the key players in this pathway, including different zinc finger proteins and WRKY transcription factors, are also central regulators of abiotic stress responses involved in temperature, salinity and osmotic stresses. Here, we describe our recent findings and discuss how ROS integrate different signals originating from different cellular compartments during abiotic stress. [source] Recent advances in the role and biosynthesis of ascorbic acid in plantsPLANT CELL & ENVIRONMENT, Issue 4 2001P. L. Conklin ABSTRACT The past few years have provided many advances in the role and biosynthesis of L -ascorbic acid (AsA) in plants. There is an increasing body of evidence confirming that AsA plays an important role in the detoxification of reactive oxygen species. The role of AsA in photoprotection has been confirmed in vivo with the use of Arabidopsis mutants. A player in the defence against reactive oxygen species, AsA peroxidase, has been extensively studied at the molecular level, and regulation of this key enzymatic activity appears to occur at several levels. As a cofactor in the hydroxylation of prolyl and lysl-residues by peptidyl-prolyl and -lysyl hydroxylases, AsA plays a part in cell wall synthesis, defence, and possibly cell division. The maintenance of reduced levels of AsA appears to be highly regulated, involving the interplay of both monodehydroascorbate and dehydroascorbate reductases and possibly auxin. A major breakthrough in plant AsA biosynthesis has been made recently, and strong biochemical and genetic evidence suggest that GDP-mannose and L -galactose are key substrates. In addition, evidence for an alternative AsA biosynthetic pathway(s) exists and awaits additional scrutiny. Finally, newly described Arabidopsis mutants deficient in AsA will further increase our understanding of AsA biosynthesis [source] Rescue of the reeler phenotype in the dentate gyrus by wild-type coculture is mediated by lipoprotein receptors for reelin and disabled 1THE JOURNAL OF COMPARATIVE NEUROLOGY, Issue 1 2006Shanting Zhao Abstract Reelin is a positional signal for the lamination of the dentate gyrus. In the reeler mutant lacking Reelin, granule cells are scattered all over the dentate gyrus. We have recently shown that the reeler phenotype of the dentate gyrus can be rescued in vitro by coculturing reeler hippocampal slices with slices from wild-type hippocampus. Here we studied whether Reelin from other brain regions can similarly induce this rescue effect and whether it is mediated via the Reelin receptors apolipoprotein E receptor 2 (ApoER2) and very-low-density lipoprotein receptor (VLDLR). We found that coculturing reeler hippocampal slices with slices from wild-type olfactory bulb, cerebellum, and neocortex rescued the reeler phenotype as seen before with hippocampal slices, provided that the Reelin-synthesizing cells of these regions were placed near the marginal zone of the reeler hippocampal slice. However, coculturing wild-type hippocampal slices with hippocampal slices from mutants deficient in ApoER2 and VLDLR did not rescue the reeler-like phenotype in these cultures. Similarly, no rescue of the reeler-like phenotype was observed in slices from mutants lacking Disabled 1 (Dab1), an adapter protein downstream of Reelin receptors. Conversely, reeler hippocampal slices were rescued by coculturing them with slices from Dab1,/, mutants or ApoER2,/,/VLDLR,/, mice. These findings show that Reelin from other brain regions can substitute for the loss of hippocampal Reelin and that rescue of the reeler phenotype observed in our coculture studies is mediated via lipoprotein receptors for Reelin and Dab1. J. Comp. Neurol. 495:1,9, 2006. © 2006 Wiley-Liss, Inc. [source] Role of EHEC O157:H7 virulence factors in the activation of intestinal epithelial cell NF-,B and MAP kinase pathways and the upregulated expression of interleukin 8CELLULAR MICROBIOLOGY, Issue 10 2002M. Cecilia Berin Summary Enterohaemorrhagic Escherichia coli O157:H7 (EHEC) is a gastrointestinal pathogen that is generally non-invasive for intestinal epithelial cells, yet causes acute intestinal inflammation, diarrhoea, haemorrhagic colitis and haemolytic uraemic syndrome. To study signal transduction pathways activated in human intestinal epithelial cells by EHEC, we took advantage of EHEC O157:H7 and isogenic mutants deficient in the major EHEC virulence factors, intimin (eae,) and Shiga toxin (stx,). Infection with wild-type EHEC activated p38 and ERK MAP kinases and the nuclear translocation of the transcription factor NF-,B. Downstream, this was accompanied by increased expression of mRNA and protein for the neutrophil chemoattractant IL-8. Isogenic eae, and stx, mutants also activated p38 and ERK MAP kinases, and NF-,B and stimulated increases in IL-8 protein secretion similar to those of wild-type EHEC. Further, inhibition of either p38, ERK or NF-,B activation abrogated the IL-8 response induced by wild-type EHEC and the mutants. Epithelial cell MAP kinase and NF-,B pathways leading to IL-8 secretion were also activated by isolated EHEC H7 flagellin, which was active when added to either the apical or basolateral surface of polarized human intestinal epithelial cells. We conclude that EHEC interacting with intestinal epithelial cells activates intracellular signalling pathways and an epithelial cell proinflammatory response independent of either Shiga toxin or intimin, two of the major known virulence factors of EHEC. The activation of proinflammatory signals in human colon epithelial cells in response to this non-invasive pathogen appears to depend to a significant extent on H7 flagellin. [source] | ||||||||||||||||||||||