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PubMed1982

Molecular cloning of sequences from a Drosophila RNA polymerase II locus by P element transposon tagging.

We have identified a lethal mutation in the D. melanogaster RNA polymerase II locus, RpIIC4, caused by insertion of a transposable element associated with the phenomenon of hybrid dysgenesis (P element). Using previously cloned P element sequences as a hybridization probe we have isolated a hybrid lambda phage clone carrying a 10 kb genomic DNA fragment containing a 1.3 kb P element insert and flanking sequences from the RpII locus. The non-P sequences in this clone (lambda DmRpII-1) hybridize to polytene chromosome band region 10C, the cytogenetic location of RpIIC4, and revertants which lose the lethal RNA polymerase II mutation also lose P element sequences from the locus. We have generated several additional P element insertions into the locus and shown that they can occur at two or more different sites. These experiments illustrate that mutagenesis by P element insertion and use of cloned P DNA to retrieve the DNA sequences into which insertion has occurred may be a general method for cloning genetically defined loci in Drosophila.

Alleles↗
PubMed1984

Introduction of a functional P element into the germ-line of Drosophila hawaiiensis.

When a plasmid carrying a P-transposable element (derived from Drosophila melanogaster) is injected into young embryos of D. hawaiiensis, the P-element sequence from the plasmid transposes into the germ-line chromosomes. The introduction of this P element into D. hawaiiensis provides an opportunity to study the behavior of the transposable element in a novel context. Germ-line transposition and numerical increase of the P elements are readily detected in D. hawaiiensis. Thus these aspects of P-element function do not require chromosomal or cytoplasmic properties that are unique to D. melanogaster. Since D. hawaiiensis is among those Drosophila that are most distantly related to D. melanogaster, these results suggest that P-element-mediated transformation may function in many species.

Animals↗
PubMed1990

Identification of an insertion-sequence-like genetic element in the newly recognized human pathogen Mycoplasma incognitus.

Cloned 2.2-kb DNA (plasmid psb-2.2) of Mycoplasma incognitus, a pathogen in AIDS and non-AIDS patients [Lo et al., Am. J. Trop. Med. Hyg. 41 (1989) 364-376; 601-616], contains a 1405-bp genetic element closely resembling bacterial insertion sequence (IS) elements. This IS-like element has 29-bp terminal inverted repeats with seven mismatches, is immediately flanked by 3-bp direct repeats, and has typical stem-and-loop structures at or near both the termini. Two potential open reading frames (ORF-1 and ORF-2) encode 143 amino acids (aa) and 103 aa, respectively, in this IS-like element. Part (57 aa) of the deduced aa sequence of ORF-2 has a significant homology (43%) with the putative transposase of Escherichia coli IS3. In this study, a series of synthetic oligodeoxyribonucleotides each containing a specific sequence of a selected segment in psb-2.2, have been used as probes which reveal that the IS-like element occurs more than ten times in the genome of M. incognitus. This potentially transposable element has many characteristic features in common with bacterial IS elements.

Amino Acid Sequence↗
PubMed1996

Vector for IS element entrapment and functional characterization based on turning on expression of distal promoterless genes.

We constructed and characterized a novel trap vector for rapid isolation of insertion sequences. The strategy used for the isolation of IS elements is based on the ability of many IS elements to turn on the expression of otherwise silent genes distal to some sites of insertion. The simple transposition of an IS element can sometimes cause the constitutive expression of promoterless antibiotic resistance genes resulting in selectable phenotypes. The trap vector pAW1326 is based on a pBR322 replicon, it carries ampicillin and streptomycin resistance genes, and also silenced genes that confer chloramphenicol and kanamycin resistance once activated. The trap vector pAW1326 proved to be efficient and 85 percent of all isolated mutations were insertions. The majority of IS elements resident in the studied Escherichia coli strains tested became trapped, namely IS2, IS3, IS5, IS150, IS186 and Tn1000. We also encountered an insertion sequence, called IS10L/R-2, which is a hybrid of the two IS variants IS10L and IS10R. IS10L/R-2 is absent from most E. coli strains, but it is detectable in some strains such as JM109 which had been submitted to Tn10 mutagenesis. The distribution of the insertion sequences within the trap region was not random. Rather, the integration of chromosomal mobile genetic elements into the offered target sequence occurred in element-specific clusters. This is explained both by the target specificity and by the specific requirements for the activation of gene transcription by the DNA rearrangement. The employed trap vector pAW1326 proved to be useful for the isolation of mobile genetic elements, for a demonstration of their transposition activity as well as for the further characterization of some of the functional parameters of transposition.

DNA Transposable Elements↗
PubMed1996

Identification and characterization of transcriptional regulatory elements of the human aromatase cytochrome P450 gene (CYP19).

Aromatase cytochrome P450, a member of the cytochrome P450 gene super family, catalyzes conversion of androgens to estrogens in a form of an enzyme-complex with NADPH-cytochrome P450 reductase. Transcription of the aromatase cytochrome P450 gene (CYP19) is regulated in part by tissue-specific promoters coupled with alternative splicing mechanisms. The transcription in human placenta is governed by a promoter activity of the 5' flanking region of exon I.1, which is mapped more than 40 kb upstream from the translational start codon observed in exon II. Transient expression analyses with chimeric constructs containing the 5' flanking sequences linked to the bacterial chloramphenicol acetyltransferase (CAT) gene in human BeWo choriocarcinoma cells localized a cell-type specific enhancer element between -242 and -166 relative to the major cap site. DNase I footprinting and transient expression analyses of the enhancer element indicate that it consists of two sub-elements and that both sub-elements are necessary for the maximum enhancement of the transcription. In addition to the enhancer element, a cis-acting element important for transcriptional enhancement of the gene in response to 12-O-tetradecanoylphorbol 13-acetate in BeWo cells is localized between -2141 and -2115. A nuclear factor binding to the element is identified as NF-IL6 (also termed as LAP and C/EBP beta). Transient expression analyses using the CAT constructs containing the NF-IL6 binding sites involvement of the factor in transcriptional regulation of CYP19.

Aromatase↗
PubMed2005

Peroxisome proliferator-activated receptor alpha and its response element are required but not sufficient for transcriptional activation of the mouse heart-type fatty acid binding protein gene.

Expression of heart-type fatty acid binding protein is restricted mainly to the skeletal and cardiac muscles and further regulated by peroxisome proliferator-activated receptor alpha. The molecular basis for the muscle-restricted peroxisome proliferator-activated receptor alpha action on the fatty acid binding gene was analyzed using normal and the receptor-null mice and the cultured cells. Two possible peroxisome proliferator-response elements were found in the promoter region of the mouse gene. A gel shift assay showed that both elements were functional. However, neither the tandem repeats of the elements nor the cloned promoter sequence could be activated by peroxisome proliferator-activated receptor alpha and its ligand in the reporter gene assay using cultured cells. The cloned promoter responded to the ligand only in the muscle when the reporter gene was introduced into the mouse muscle. Using a chimeric receptor with the activation domain of herpes virus VP16 protein and the tandem repeats of the elements with or without mutation, the upstream element was finally demonstrated to be potentially involved in the receptor-dependent transcriptional activation. These results suggest that the peroxisome proliferator-response element of the mouse gene is atypical and there is a muscle-specific mechanism to enhance the weak binding of the receptor to the response element to ensure the muscle-specific action of peroxisome proliferator-activated receptor alpha on the heart-type fatty acid binding protein gene promoter.

Animals↗
PubMed2006

Micro-chemical imaging of cesium distribution in Arabidopsis thaliana plant and its interaction with potassium and essential trace elements.

Cesium as an alkali element exhibits a chemical reactivity similar to that of potassium, an essential element for plants. It has been suggested that Cs phytotoxicity might be due either to its competition with potassium to enter the plant, resulting in K starvation, or to its intracellular competition with K binding sites in cells. Such elemental interactions can be evidenced by chemical imaging, which determines the elemental distributions. In this study, the model plant Arabidopsis thaliana was exposed to 1 mM cesium in the presence (20 mM) or not of potassium. The quantitative imaging of Cs and endogenous elements (P, S, Cl, K, Ca, Mn, Fe, and Zn) was carried out using ion beam micro-chemical imaging with 5 microm spatial resolution. Chemical imaging was also evidenced by microfocused synchrotron-based X-ray fluorescence (microXRF) which presents a better lateral resolution (<1 microm) but is not quantitative. Cesium distribution was similar to potassium which suggests that Cs can compete with K binding sites in cells. Cesium and potassium were mainly concentrated in the vascular system of stems and leaves. Cs was also found in lower concentration in leaves mesophyll/epidermis. This late representing the larger proportion in mass, mesophyll/epidermis can be considered as the major storage site for cesium in A. thaliana. Trichomes were not found to accumulate cesium. Interestingly, increased Mn, Fe, and Zn concentrations were observed in leaves at high chlorosis. Mn and Fe increased more in the mesophyll than in veins, whereas zinc increased more in veins than in the mesophyll suggesting a tissue specific interaction of Cs with these trace elements homeostasis. This study illustrates the sensitivity of ion beam microprobe and microfocused synchrotron-based X-ray fluorescence to investigate concentrations and distributions of major and trace elements in plants. It also shows the suitability of these analytical imaging techniques to complement biochemical investigations of metallic stress in plants.

Arabidopsis↗
PubMed2005

Biomonitoring of 30 trace elements in urine of children and adults by ICP-MS.

The paper provides physicians and clinical chemists with statistical data (concentration ranges, geometric mean values, selected percentiles, etc.) about 30 urinary trace elements in order to determine whether people have trace element deficiencies or have been exposed to higher elemental concentrations. Morning urine samples of 72 children and 87 adults from two geographical areas of Germany were collected and the elements Li, Be, V, Cr, Mn, Ni, Co, Cu, Zn, Ga, As, Se, Rb, Sr, Mo, Rh, Pd, Ag, Cd, In, Sn, Sb, Cs, Ba, Pt, Au, Pb, Tl, Bi and U were determined by inductively coupled plasma mass spectrometry (ICP-MS) with a new octopole based collision/reaction cell. The urine samples were analysed directly after a simple 1/5 (V/V) dilution with deionised water and nitric acid. Information on exposure conditions of all human subjects were collected by questionnaire-based interviews. The described concentration data down to the ng/l range are very useful for the formulation of reference values. For some elements either new data are described (e.g., for V, Ga, In, Bi, Rh, Mn) or differences to earlier studies were found (e.g., for Be, As). For other elements (e.g., Sb, Se, Mo, Ba, Cu, Zn, Li) our results are in good correlation with previous studies and also complemented with urinary trace element concentrations for children.

Adult↗
PubMed2005

Trace element accumulation in the moss Hypnum cupressiforme Hedw. and the trees Quercus ilex L. and Pinus halepensis Mill. in Catalonia.

We studied trace element accumulation in the moss Hypnum cupressiforme and the widely distributed Mediterranean trees Quercus ilex and Pinus halepensis located at increasing distances from the Barcelona Metropolitan Area. Hypnum cupressiforme, Quercus ilex and, to a somewhat lesser extent, Pinus halepensis, have proved to be adequate as possible accumulative monitoring species in relation to trace elements pollution. No significant effects of crown orientation were found. One-year old leaves generally accumulated more trace elements than current-year leaves. All the studied trace elements showed greatest concentrations in the Barcelona Metropolitan Area, with lead, cadmium and arsenic concentrations being especially high. In general, trace element concentrations in biomass were similar or higher than the values reported from other Mediterranean urban areas of Europe. The top soil-layer concentrations were also higher in the Barcelona Metropolitan Area indicating the existence of mechanisms of atmospheric deposition and/or concentration in the soil. The lower values of Pb of airborne origin relative to other elements such as Cd, Cu, Zn and Sb suggest that traffic exhausts are not the only important focus of pollutants in this area. The results of biomass concentrations and of enrichment factor of biomasses respect to bedrock and soils show that atmospheric inputs account for the higher trace element concentrations in the Barcelona Metropolitan Area.

Biomass↗
PubMed2006

Trace elements in atmospheric particulate matter over a coal burning power production area of western Macedonia, Greece.

Total suspended particle (TSP) concentrations were determined in the Eordea basin (western Macedonia, Greece), an area with intensive lignite burning for power generation. The study was conducted over a one-year period (November 2000-November 2001) at 10 sites located at variable distances from the power plants. Ambient TSP samples were analyzed for 27 major, minor and trace elements. Annual means of TSP concentrations ranged between 47+/-33 microg m(-3) and 110+/-50 microg m(-3) at 9 out of the 10 sites. Only the site closest to the power stations and the lignite conveyor belts exhibited annual TSP levels (210+/-97 microg m(-3)) exceeding the European standard (150 microg m(-3), 80/779/EEC). Concentrations of TSP and almost all elemental components exhibited significant spatial variations; however, the elemental profiles of TSP were quite similar among all sites suggesting that they are affected by similar source types. At all sites, statistical analysis indicated insignificant (P<0.05) seasonal variation for TSP concentrations. Some elements (Cl, As, Pb, Br, Se, S, Cd) exhibited significantly higher concentrations at certain sites during the cold period suggesting more intense emissions from traffic, domestic heating and other combustion sources. On the contrary, concentrations significantly higher in the warm period were found at other sites mainly for crustal elements (Ti, Mn, K, P, Cr, etc.) suggesting stronger influence from soil resuspension and/or fly ash in the warm months. The most enriched elements against local soil or road dust were S, Cl, Cu, As, Se, Br, Cd and Pb, whereas negligible enrichment was found for Ti, Mn, Mg, Al, Si, P, Cr. At most sites, highest concentrations of TSP and elemental components were associated with low- to moderate-speed winds favoring accumulation of emissions from local sources. Influences from the power generation were likely at those sites located closest to the power plants and mining activities.

Air Pollutants↗
PubMed2006

A comparison of phytoremediation capability of selected plant species for given trace elements.

In our experiment, As, Cd, Pb, and Zn remediation possibilities on medium contaminated soil were investigated. Seven plant species with a different trace element accumulation capacity and remediation potential were compared. We found good accumulation capabilities and remediation effectiveness of Salix dasyclados similar to studied hyperaccumulators (Arabidopsis halleri and Thlaspi caerulescens). We have noticed better remediation capability in willow compared to poplar for most of the elements considered in this experiment. On the contrary, poplar species were able to remove a larger portion of Pb as opposed to other species. Nevertheless, the removed volume was very small. The elements found in plant biomass depend substantially on the availability of these elements in the soil. Different element concentrations were determined in natural soil solution and by inorganic salt solution extraction (0.01 molL(-1) CaCl(2)). Extracted content almost exceeded the element concentration in the soil solution. Element concentrations in soil solution were not significantly affected by sampling time.

Arabidopsis↗
PubMed2004

Analysis of essential elements in Pragya-peya--a herbal drink and its constituents by neutron activation.

Ayurvedic herbal formulations are a good source of several nutrient elements essential for metabolic processes. Pragya-peya, a herbal drink and its 12 herbal constituents have been analyzed for 7 minor (Al, Ca, Cl, Mg, Na, K, P) and 15 trace (Ba, Br, Co, Cr, Cs, Fe, Hg, La, Mn, Rb, Sc, Se, Th, V, Zn) elements by instrumental neutron activation analysis (INAA). The method involves thermal neutron irradiation in a reactor followed by counting at several intervals. Also Cd, Cu, Ni and Pb contents were determined by atomic absorption spectrometry (AAS). Elemental data were validated by simultaneously analysing reference materials (RMs) such as mixed Polish herbs (INCT-MPH-2) and peach leaves (SRM-1547). Sample homogeneity was tested by analysing samples from three different batches collected at different intervals. Pragya-peya has been found to be especially rich in several nutrient elements such as Ca, K, V, Fe, Mn, Se and Zn whereas no single constituent is enriched in all the nutrient elements. Concentrations of elements are discussed vis-a-vis their medicinal/therapeutic uses. Several elements such as Na, K, P, Ca, Fe, Co, and Zn seem to be well correlated in 12 constituent herbs.

Beverages↗
PubMed2005

GAAAATATGATA-like elements in androgen-associated regulation of the prostatic acid phosphatase gene.

Purpose of the study was to clarify molecular mechanisms behind tissue-specific and hormone-dependent gene expression using human prostatic acid phosphatase (hPAP) gene as a model. Regulatory region -734/+467 of hPAP gene induces transcription of a reporter gene in the prostate of transgenic mice. It contains five elements, A-E, homologous to GAAAATATGATA sequence, which is connected to prostate-specific and androgen-dependent gene expression. The significance of the C, D and E elements in the transcriptional regulation of hPAP gene was evaluated using reporter gene assays. The deletion of element C from the hPAP promoter constructs mainly decreased their transcriptional activity in the presence of androgen, while increased activity particularly in the absence of androgens was detected after removal of elements D and E. These events took place in transiently transfected prostatic LNCaP cells, but not in non-prostatic COS-1 cells. As a conclusion, the GAAAATATGATA-like elements are involved in the transcriptional regulation of hPAP promoter constructs in prostatic cells. These elements mediate both transcriptional activation and repression depending on the hormonal status of the cells and location of the element in the construct.

Acid Phosphatase↗
PubMed2005

The role of DNA response elements as allosteric modulators of steroid receptor function.

Steroid receptors are ligand-activated transcription factors which control the expression of their target genes by binding to specific DNA elements. Consensus response elements have been delineated for the glucocorticoid, androgen, progesterone and mineralocorticoid receptors on one hand (steroid response element, SRE) and for the estrogen receptor on the other hand (estrogen response element, ERE). Small variations in these sequences not only affect the binding but may also have a dramatic impact on the transcriptional activity of steroid receptors. It has now become obvious that DNA response elements do not merely tether regulatory proteins to control regions of target genes but may additionally impart conformational changes onto the DNA-binding domain as well as to neighbouring domains of steroid receptors. This in turn will create unique platforms for selective recruitment of cofactors and possibly for induction of modifications in local chromatin architecture. An additional level of complexity is added by the frequent presence of multiple response elements in gene promoter regions. The allosteric effects of DNA response elements on steroid receptors may be essential for differential gene expression and this offers interesting perspectives for the identification of selective modulators.

Allosteric Regulation↗
PubMed2005

Temporal trends of total and potentially available element concentrations in sewage biosolids: a comparison of biosolid surveys conducted 18 years apart.

The application of biosolids to agricultural land is becoming increasingly popular because of the many benefits they can offer in terms of improved soil fertility and productivity, and also because land application offers an economically viable disposal option. However, the heavy metal and nutrient content of biosolids are potential causes for concern due to the possibility of their accumulation in soils to levels that may adversely affect the environment. Therefore, total concentrations of elements in biosolids and the fraction potentially available to organisms must be known in order to accurately gauge the risks and to classify the biosolids. In 1983, deVries conducted a survey to determine the total and available contents of elements in biosolids collected from various treatment plants around Australia. We conducted a similar survey on biosolids collected in 2001, and thus were able to identify temporal trends in biosolid characteristics. The 2001 survey included biosolids from several treatment plants sampled in 1983, thus some direct comparisons were possible, and in four cases, it was possible to examine temporal trends in biosolid characteristics within individual treatment plants. Total element concentrations were determined by aqua regia digestion, and available element concentrations were determined by 0.01 M CaCl(2) extraction. Results showed total concentrations of Cu, Mn, Ni, Na, and Ca changed little over the period examined, however the 2001 median values of Cd, Mg, Pb, and Zn were all reduced by 60% or greater. Median total concentration of K was more than 50% higher, and P 120% higher, in the 2001 survey. Changes in the availability of elements surveyed in 1983 and 2001 were varied for different elements and for different biosolids. Extractable element concentrations (viewed as percentages of total concentrations) of Cu, Cd, and Ni all fell by 50-72% between 1983 and 2001, while K extractability fell by 35%.

Agriculture↗
PubMed2006

Trace element availability and plant growth in a mine-spill-contaminated soil under assisted natural remediation II. Plants.

In this second part, we evaluated the effects of different amendments on plant growth (Agrostis stolonifera L.), and trace element accumulation and removal by plants in a trace element (As, Cd, Cu, Pb and Zn) contaminated soil. Description of the various treatments is given in Part I of this work. The plants were grown for consecutive periods (2002, 2003, 2004), 5 months each and harvested twice in each period. Results showed that plant growth was enhanced and trace element concentrations in plant were reduced in SL, MWC, BC and LEO treatments in the first period. No significant differences were observed in subsequent periods. This seemed to be related with changes in soil pH. Removal of trace elements was higher in SL, MWC, BC and LEO treatments due to higher biomass production in the first period. In following years no significant differences between treatments were found. Data from Part I of this study were also used to compare trace element bioavailable concentrations extracted with 0.01 M CaCl2 and 0.05 M EDTA with trace elements in plant. We observed that 0.01 M CaCl2 was more suitable for determination of bioavailable concentrations and that extraction with EDTA overestimated biovailability of trace elements in amended treatments, especially in those where composts were added.

Agrostis↗
PubMed2004

An internal element of the measles virus antigenome promoter modulates replication efficiency.

The cis-acting sequence elements that direct measles virus (MV) genome synthesis reside in the 109 base non-coding region at the 5' trailer (3' antigenomic) end of MV genome that makes up the antigenomic promoter (AGP). The MV-AGP nucleotides 79-96, corresponding to nucleotide hexamers 14, 15 and 16 (the C' element), show sequence similarity with the equivalent region of many paramyxoviruses and are analogous to the three nucleotide hexamers that form the second replication control element in the Sendai virus AGP. In this study, results of two independent procedures demonstrate that the MV C' element also is a replication control sequence. Results of in vivo nucleotide selection experiments show that selection pressure for retaining the wild type nucleotides at the first position of each of the three hexamers, and for the fifth position of the 14th hexamer was relatively high. However, with continued replication, preference for the conservation of wild type nucleotides across the entire C' element was clearly evident. Results of mutational analysis of individual nucleotides in one or more hexamers in a measles-helper-virus driven reporter gene rescue system agreed with these results. Substitutions at the first position of the 14th, the 15th or the 16th hexamers reduced minireplicon activity dramatically. In contrast, changes at the other five positions of any one hexamer had little or no effect on minireplicon activity, even when all the five bases were changed at the same time. However, when minireplicons were analyzed which contained point mutations at equivalent positions in all three hexamers, it was evident that the nucleotides, particularly those at the 5th position, were also important components of the C' element. This pattern of sequence requirement in the C' element based on mutational analysis could be described as a distinct motif, 5'-(GNNNAN)2GNNNCN-3', that is important for MV replication.

Base Sequence↗
PubMed2000

Toxic and essential elements in placentas of Swedish women.

OBJECTIVES: To evaluate interactions between toxic and essential elements in the mother-fetus relationship and possible predictors of trace element concentrations in placenta and cord blood. DESIGN AND METHODS: A group of 106 Swedish women was investigated for concentrations of cadmium, lead, and several essential elements in placenta as well as cadmium, lead, zinc, and selenium in venous blood collected at gestational week (gw) 36 and umbilical cord blood. Relations between these elements and maternal and child's characteristics were examined. RESULTS: The concentrations of cadmium in placenta ranged from 10 to 170 nmol/kg, with the median value (Md) being 46 nmol/kg. Cord blood cadmium (Md of 0.19 nmol/L) was only about 10% of that in maternal blood. Smokers had significantly higher cadmium concentrations in blood (p < 0.001) and placenta (p = 0.001) than non-smokers. The median placental concentration of lead was 26 nmol/kg (range 0-630 nmol/kg). The lead levels in cord blood (Md of 54 nmol/L) were almost the same as in maternal blood. Statistically significant negative associations were found between cord blood lead, on one hand, and child's weight, length, and head circumference, on the other. The placental levels (medians and ranges) of the essential elements (micromol/kg) were 160 (120-280) for zinc, 2.4 (2.0-3.3) for selenium, 15 (10-20) for copper, 0.084 (0.02-0.32) for cobalt, 0.055 (0.03-0.12) for molybdenum, and 1.2 (0. 65-5.1) for manganese, respectively. Several of the essential elements in placenta correlated significantly with each other. Multiparous mothers had significantly lower concentrations of zinc (p = 0.002) and selenium (p = 0.049) in serum as well as zinc (p = 0. 001) and calcium (p = 0.004) in placenta than nulliparous ones. Also, cord blood zinc decreased with parity. CONCLUSIONS: The results showed that lead, but not cadmium crossed easily the placental barrier. There were no negative effects of cadmium on the zinc status. Cord blood lead, on the other hand, was a negative predictor of child's birth weight, length and head circumference, indicating that lead might have negative influence on growth in children even at very low exposure levels. There was a depletion of maternal stores of essential elements with increasing parity.

Cadmium↗