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1.
Transcriptional control, translation and function of the products of the five open reading frames of the Escherichia coli nir operon 总被引:3,自引:0,他引:3
Nerina R. Harborne Lesley Griffiths Stephen J. W. Busby Jeffrey A. Cole 《Molecular microbiology》1992,6(19):2805-2813
Five open reading frames designated nirB, nirD, nirE, nirC and cysG have been identified from the DNA sequence of the Escherichia coli nir operon. Complementation experiments established that the NirB, NirD and CysG polypeptides are essential and sufficient for NADH-dependent nitrite reductase activity (EC 1.6.6.4). A series of plasmids has been constructed in which each of the open reading frames has been fused in-phase with the beta-galactosidase gene, lacZ. Rates of beta-galactosidase synthesis during growth in different media revealed that nirB, -D, -E and -C are transcribed from the FNR-dependent promoter, p-nirB, located just upstream of the nirB gene: expression is co-ordinately repressed by oxygen and induced during anaerobic growth. Although the nirB, -D and -C open reading frames are translated into protein, no translation of nirE mRNA was detected. The cysG gene product is expressed from both p-nirB and a second, FNR-independent promoter, p-cysG, located within the nirC gene. No NADH-dependent nitrite reductase activity was detected in extracts from bacteria lacking either NirB or NirD, but a mixture of the two was as active as an extract from wild-type bacteria. Reconstitution of enzyme activity in vitro required stoichiometric quantities of NirB and NirD and was rapid and independent of the temperature during mixing. NirD remained associated with NirB during the initial stages of purification of the active enzyme, suggesting that NirD is a second structural subunit of the enzyme. 相似文献
2.
M N Berry R B Gregory A R Grivell D C Henly C D Nobes J W Phillips P G Wallace 《Biochimica et biophysica acta》1988,936(3):294-306
The lipophilic triphenylmethylphosphonium cation (TPMP+) has been employed to measure delta psi m, the electrical potential across the inner membrane of the mitochondria of intact hepatocytes. The present studies have examined the validity of this technique in hepatocytes exposed to graded concentrations of inhibitors of mitochondrial energy transduction. Under these conditions, TPMP+ uptake allows a reliable measure of delta psi m in intracellular mitochondria, provided that the ratio [TPMP+]i/[TPMP+]e is greater than 50:1 and that at the end of the incubation more than 80% of the hepatocytes exclude Trypan blue. Hepatocytes, staining with Trypan blue, incubated in the presence of Ca2+, do not concentrate TPMP+. The relationships between delta psi m and two other indicators of cellular energy state, delta GPc and Eh, or between delta psi m and J0, were examined in hepatocytes from fasted rats by titration with graded concentrations of inhibitors of mitochondrial energy transduction. Linear relationships were generally observed between delta psi m and delta GPc, Eh or J0 over the delta psi m range of 120-160 mV, except in the presence of carboxyatractyloside or oligomycin, where delta psi m remained constant. Both the magnitude and the direction of the slope of the observed relationships depended upon the nature of the inhibitor. Hepatocytes from fasted rats synthesized glucose from lactate or fructose, and urea from ammonia, at rates which were generally linear functions of the magnitude of delta psi m, except in the presence of oligomycin or carboxyatractyloside. Linear relationships were also observed between delta psi m and the rate of formation of lactate in cells incubated with fructose and in hepatocytes from fed rats. The linear property of these force-flow relationships is taken as evidence for the operation of thermodynamic regulatory mechanisms within hepatocytes. 相似文献
3.
Summary Worldwide, invasive weeds threaten agricultural, natural and urban ecosystems. In Australia's agricultural and grazing regions, invasive species often establish across extensive areas where weed management is hampered by an inability to detect the location and timing of an outbreak. In these vast landscapes, an effective detection and monitoring system is required to delineate the extent of the invasion and identify spatial and temporal factors associated with weed establishment and thickening. In this study, we utilize a time series of remote sensing imagery to detect the spatial and temporal patterns of Prickly Acacia ( Acacia nilotica ) invasion in the Mitchell grass plains of North Queensland. We develop a spectral index from Landsat images which is applied to images from 1989 to 2004, in combination with a classification mask, to identify locations and monitor changes in Prickly Acacia density across 29 000 km2 of Mitchell grass plains. The approach identified spectral and temporal signatures consistent with Prickly Acacia infestation on 1.9% of this landscape. Field checking of results confirmed presence of the weed in previously unrecorded locations. The approach may be used to evaluate future spread, or outcomes of management strategies for Prickly Acacia in this landscape and could be employed to detect and monitor invasions in other extensive landscapes. 相似文献
4.
Rachael V. Gallagher Lesley Hughes Michelle R. Leishman Peter D. Wilson 《Biological invasions》2010,12(12):4049-4063
Potential interactions between climate change and exotic plant invasions may affect areas of high conservation value, such
as land set aside for the protection of endangered species or ecological communities. We investigated this issue in eastern
Australia using species distribution models for five exotic vines under climate regimes for 2020 and 2050. We examined how
projected changes in the distribution of climatically suitable habitat may coincide with the remaining remnants of an endangered
ecological community—littoral rainforests—in this region. The number of known infestations of each weed in tropical, subtropical
and temperate areas was used to assess the likelihood of further expansion into areas projected to provide suitable habitat
under future conditions. Littoral rainforest reserves were consistently predicted to provide bioclimatically suitable habitat
for the five vines examined under both current and future climate scenarios. We explore the consequences and potential strategies
for managing exotic plant invasions in these protected areas in the coming decades. 相似文献
5.
T J Wallace E M Kodsi T B Langston M R Gergis W M Grogan 《The Journal of biological chemistry》2001,276(35):33165-33174
Site-directed mutagenesis is used to identify amino acid residues that dictate reported differences in substrate specificity between rat hepatic neutral cytosolic cholesteryl ester hydrolase (hncCEH) and rat lung carboxylesterase (LCE), proteins differing by only 4 residues in their primary sequences. Beginning with LCE, the substitution Met(423) --> Ile(423) alone or in combination with other mutations increased activity with p-nitrophenylcaprylate (PNPC) relative to more hydrophilic p-nitrophenylacetate (PNPA), typical of hncCEH. The substitution Thr(444) --> Met(444) was necessary but not sufficient for expression of cholesteryl esterase activity in COS-7 cells. The substitution Asn(506) --> Ser(506), creating a potential phosphorylation site, uniformly increased activity with both PNPA and PNPC, was necessary but not sufficient for expression of cholesteryl esterase activity and conferred susceptibility to activation by cAMP-dependent protein kinase, a property of hncCEH. The 3 mutations in combination were necessary and sufficient for expression of cholesteryl esterase activity by the mutated LCE. The substitution Gln(186) --> Arg(186) selectively reduced esterase activity with PNPA and PNPC but was not required for cholesteryl esterase activity. Homology modeling from x-ray structures of acetylcholinesterases is used to propose three-dimensional models for hncCEH and LCE that provide insight into the effects of these mutations on substrate specificity. 相似文献
6.
Summary Detopped root systems of tobacco plants (Nicotiana tabacum L. var. Virginia Gold) were exposed to air at different temperatures (14 to 23°C). The volume of xylem exudate was related
to temperature and decreasing the temperature resulted in a lag in the appearance of exudate. For 4 and 8 days of exudation
the amounts of cations transported in the exudate were temperature dependent. The Q10 values were higher than usually encountered in biological reactions; the mean value was around 6.
This work was supported in part by National Science Foundation Grant GB8170.
University of California 相似文献
7.
William Wallace 《BMJ (Clinical research ed.)》1914,2(2795):207-208
8.
9.
G. K. Wallace 《Biological cybernetics》1989,61(3):233-240
The patterns of EMG activity in the biceps and triceps muscles were recorded during horizontal oscillatory movements of the forearm. Subjects showed increased frequency of oscillation as they voluntarily reduced movement amplitude. EMG burst duration was significantly correlated with wavelength of oscillation in every case. In almost half the cases burst intensity was also positively correlated with wavelength. Subjects seemed to be using one or both these methods to control amplitude. A model was developed in three stages which satisfactorily accounted for the data. 相似文献
10.
Caroline van Haaften-Day Peter Russell Susan Carr Lesley Wright 《In vitro cellular & developmental biology. Plant》1988,24(10):965-971
Summary A cell line derived from a human ovarian carcinosarcoma was established in tissue culture and in nude mice. Two sublines,
LDF and HDF, separated by discontinuous density centrifugation were also established from the parent line JoN. The cloning
efficiency of the JoN line was 21%. Morphologic features of adenocarcinoma cells characteristic of the parent JoN cells were
retained in the sublines and clones; all lines showed the same karyotype and DNA content (pseudodiploid and pseudotetraploid).
Keratin, as demonstrated immunohistochemically, was strongly expressed in the parent line JoN and the xenograft tumor, but
not at all in the LDF sublines and only moderately in the HDF sublines. Vimentin, however, was expressed in neither the parent
line JoN nor the xenograft tumor, but was present in both sublines. Transglutaminase and plasminogen activator activity was
high in the parent line JoN. Neither, sublines nor clones showed the same high enzyme activity as the parent line. It is concluded
that this human tumor line JoN is comprised of epithelial cells, capable of multidirectional differentiation. 相似文献