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J C Hansen

Publications and source records attributed to J C Hansen.

At least 19 recordsLinked to original sources
PubMed2000

The core histone N termini function independently of linker histones during chromatin condensation.

The relationships between the core histone N termini and linker histones during chromatin assembly and salt-dependent chromatin condensation were investigated using defined chromatin model systems reconstituted from tandemly repeated 5 S rDNA, histone H5, and either native "intact" core histone octamers or "tailless" histone octamers lacking their N-terminal domains. Nuclease digestion and sedimentation studies indicate that H5 binding and the resulting constraint of entering and exiting nucleosomal DNA occur to the same extent in both tailless and intact chromatin arrays. However, despite possessing a normal chromatosomal structure, tailless chromatin arrays can neither condense into extensively folded structures nor cooperatively oligomerize in MgCl(2). Tailless nucleosomal arrays lacking linker histones also are unable to either fold extensively or oligomerize, demonstrating that the core histone N termini perform the same functions during salt-dependent condensation regardless of whether linker histones are components of the array. Our results further indicate that disruption of core histone N termini function in vitro allows a linker histone-containing chromatin fiber to exist in a decondensed state under conditions that normally would promote extensive fiber condensation. These findings have key implications for both the mechanism of chromatin condensation, and the regulation of genomic function by chromatin.

Chromatin↗
PubMed2000

Analytical sedimentation of the IIAChb and IIBChb proteins of the Escherichia coli N,N'-diacetylchitobiose phosphotransferase system. Demonstration of a model phosphotransfer transition state complex.

The phosphoenolpyruvate:glycose transferase system (PTS) is a prototypic signaling system responsible for the vectorial uptake and phosphorylation of carbohydrate substrates. The accompanying papers describe the proteins and product of the Escherichia coli N, N-diacetylchitobiose ((GlcNAc)(2)) PTS-mediated permease. Unlike most PTS transporters, the Chb system is composed of two soluble proteins, IIA(Chb) and IIB(Chb), and one transmembrane receptor (IIC(Chb)). The oligomeric states of PTS permease proteins and phosphoproteins have been difficult to determine. Using analytical ultracentrifugation, both dephospho and phosphorylated IIA(Chb) are shown to exist as stable dimers, whereas IIB(Chb), phospho-IIB(Chb) and the mutant Cys10SerIIB(Chb) are monomers. The mutant protein Cys10SerIIB(Chb) is unable to accept phosphate from phospho-IIA(Chb) but forms a stable higher order complex with phospho-IIA(Chb) (but not with dephospho-IIA(Chb)). The stoichiometry of proteins in the purified complex was determined to be 1:1, indicating that two molecules of Cys10SerIIB(Chb) are associated with one phospho-IIA(Chb) dimer in the complex. The complex appears to be a transition state analogue in the phosphotransfer reaction between the proteins. A model is presented that describes the concerted assembly and disassembly of IIA(Chb)-IIB(Chb) complexes contingent on phosphorylation-dependent conformational changes, especially of IIA(Chb).

Amino Acid Substitution↗
PubMed2000

The yeast histone acetyltransferase A2 complex, but not free Gcn5p, binds stably to nucleosomal arrays.

We have investigated the structural basis for the differential catalytic function of the yeast Gcn5p-containing histone acetyltransferase (HAT) A2 complex and free recombinant yeast Gcn5p (rGcn5p). HAT A2 is shown to be a unique complex that contains Gcn5p, Ada2p, and Ada3p, but not proteins specific to other related HAT A complexes, e.g. ADA, SAGA. Nevertheless, HAT A2 produces the same unique polyacetylation pattern of nucleosomal substrates reported previously for ADA and SAGA, demonstrating that proteins specific to the ADA and SAGA complexes do not influence the enzymatic activity of Gcn5p within the HAT A2 complex. To investigate the role of substrate interactions in the differential behavior of free and complexed Gcn5p, sucrose density gradient centrifugation was used to characterize the binding of HAT A2 and free rGcn5p to intact and trypsinized nucleosomal arrays, H3/H4 tetramer arrays, and nucleosome core particles. We find that HAT A2 forms stable complexes with all nucleosomal substrates tested. In distinct contrast, rGcn5p does not interact stably with nucleosomal arrays, despite being able to specifically monoacetylate the H3 N terminus of nucleosomal substrates. Our data suggest that the ability of the HAT A2 complex to bind stably to nucleosomal arrays is functionally related to both local and global acetylation by the complexed and free forms of Gcn5p.

Acetyl Coenzyme A↗
PubMed2000

X-ray crystallographic and analytical ultracentrifugation analyses of truncated and full-length yeast copper chaperones for SOD (LYS7): a dimer-dimer model of LYS7-SOD association and copper delivery.

Copper-zinc superoxide dismutase (CuZnSOD) acquires its catalytic copper ion through interaction with another polypeptide termed the copper chaperone for SOD. Here, we combine X-ray crystallographic and analytical ultracentrifugation methods to characterize rigorously both truncated and full-length forms of apo-LYS7, the yeast copper chaperone for SOD. The 1.55 A crystal structure of LYS7 domain 2 alone (L7D2) was determined by multiple-isomorphous replacement (MIR) methods. The monomeric structure reveals an eight-stranded Greek key beta-barrel similar to that found in yeast CuZnSOD, but it is substantially elongated at one end where the loop regions of the beta-barrel come together to bind a calcium ion. In agreement with the crystal structure, sedimentation velocity experiments indicate that L7D2 is monomeric in solution under all conditions and concentrations that were tested. In contrast, sedimentation velocity and sedimentation equilibrium experiments show that full-length apo-LYS7 exists in a monomer-dimer equilibrium under nonreducing conditions. This equilibrium is shifted toward the dimer by approximately 1 order of magnitude in the presence of phosphate anion. Although the basis for the specificity of the LYS7-SOD interaction as well as the exact mechanism of copper insertion into SOD is unknown, it has been suggested that a monomer of LYS7 and a monomer of SOD may associate to form a heterodimer via L7D2. The data presented here, however, taken together with previously published crystallographic and analytical gel filtration data on full-length LYS7, suggest an alternative model wherein a dimer of LYS7 interacts with a dimer of yeast CuZnSOD. The advantages of the dimer-dimer model over the heterodimer model are enumerated.

Computer Simulation↗
PubMed2000

Environmental contaminants and human health in the Arctic.

Monitoring of human exposure to persistent organic pollutants POPs and heavy metals have taken place since 1994 in a circumpolar study conducted under the Arctic monitoring and assessment programme (AMAP). The study has confirmed that Arctic populations relying on marine food (Inuit) have an exposure level of POPs and methyl mercury related to the amount of traditional food eaten and in some areas at a level where health effects cannot be excluded. Weighing the benefits of traditional food against the risks, AMAP have so far recommended that consumption of traditional food continues. There is a need for dietary advice to Arctic peoples so they can make informed choices concerning the food they eat.

Animals↗
PubMed2000

Organochlorines and heavy metals in pregnant women from the Disko Bay area in Greenland.

Recent studies from Greenland and the Canadian Arctic have shown high concentrations of heavy metals, such as mercury, and organochlorines in the blood and fatty tissue of the Inuit. This is attributed in particular to their high consumption of the meat and blubber of marine mammals. In the present study, 180 pregnant women and 178 newborn babies were studied, amounting to 36% of the total number of births in the Disko Bay area during 1994-1996. The pesticides found in the highest concentrations in maternal blood were DDE (4.8 micrograms/l wet wt.), trans-nonachlor (1.6 micrograms/l) and hexachlorobenzene (1.2 micrograms/l) while the total concentration of PCB (Aroclor 1260) was 19.1 micrograms/l. Calculated on a lipid basis, concentrations were slightly higher in maternal than in cord blood. The mercury concentrations were 16.8 micrograms/l in maternal blood and 35.6 micrograms/l in cord blood. In a linear regression analysis, the concentrations of organochlorines, mercury and selenium increased with maternal age. Concentrations of mercury and cadmium increased with the consumption of marine mammals, and cadmium was associated with smoking. The contaminants are potentially toxic for several organ systems but the high concentrations of pollutants have so far not been shown to influence health in Greenland.

Adult↗
PubMed2000

Comparison of contaminants from different trophic levels and ecosystems.

The present paper provides an overview of the priority contaminants and media from the Greenland part of the Arctic Monitoring and Assessment Program. Levels and accumulation patterns of heavy metals, POPs and a radionuclide (137Cs) are compared from the terrestrial, freshwater and marine ecosystems. Of the nine compounds presented, seven (Cd, Hg, Se, sigma PCB, sigma DDT, sigma HCH, HCB) increased in concentration towards higher trophic levels. For these contaminants the concentrations in soil and aquatic sediment were in the same order of magnitude, whereas the concentrations in marine biota were higher than found in the freshwater and terrestrial ecosystems probably due to the presence of longer food chains. Pb and 137Cs showed the reverse pattern compared with the other compounds. The concentrations in soil and aquatic sediments decreased in the order terrestrial, freshwater and marine ecosystems, which was reflected in the biota as well. Reindeer had similar or lower levels of Pb and 137Cs than lichens. Levels of Pb and 137Cs in marine biota did not show the same clear increase towards higher trophic as found for the other analysed compounds. Greenland Inuit contains considerably less mercury but higher levels of sigma PCB, sigma DDT and HCB than other Arctic marine top consumers.

Animals↗
PubMed2000

Elements in autopsy liver tissue samples from Greenlandic Inuit and Danes. II. Iron measured by X-ray fluorescence spectrometry.

The purpose of this study was to measure the content of iron (Fe) in liver tissue samples from urbanized Greenlandic Inuit using X-ray fluorescence spectrometry, and compare the results with those obtained in liver tissue samples from urbanized Danes. Normal liver tissue samples were obtained at autopsy from 50 Greenlandic Inuit (27 men, 23 women) with a median age of 61 years (range 23-83) and from 72 Danes (42 men, 30 women) with a median age of 62 years (range 15-87). In the entire series, there was no significant difference between liver iron in Inuit compared with Danes. Likewise, there was no significant gender difference concerning liver iron content, either in Inuit or in Danes. The median iron content (with 5-95 percentile) in Inuit was 17.23 mmol/kg dry liver (5.80-91.80) and in Danes 16.51 mmol/kg dry liver (7.83-39.05). However, when stratified according to age, a trend was revealed showing that Inuit men and women < or = 50 years had a lower liver iron content than Danes (p = 0.05 and p = 0.08) whereas Inuit men and women > 50 years had a higher liver iron content than Danes (p = 0.18 and p = 0.02). There was a significant correlation between liver iron content and age in both Inuit men (rs = 0.49, p = 0.01) and in women (rs = 0.64, p = 0.003), but not in Danes. In Inuit, the median hepatic iron index (liver iron content divided by age) was 0.33 in men and 0.32 in women. The median estimated iron content in the whole liver was 6.54 mmol (365 mg) in Inuit men and 5.41 mmol (302 mg) in Inuit women (p = 0.6). There was no correlation between hepatic iron index and age. In Danes, the median hepatic iron index was 0.46 in men and 0.29 in women (p = 0.01). There was a significant inverse correlation between hepatic iron index and age in the two genders and in the entire series (rs = -0.71, p = 0.0001). The results indicate that young and middle-aged urbanized Inuit have slightly smaller iron stores than urbanized Danes, whereas elderly Inuit have higher iron stores than Danes. In Danes, iron stores plateau at 30 to 40 years of age in men and some years after the menopause in women. In Inuit, iron stores continue to increase in old age, probably to due a lifelong dietary intake of haem iron.

Adolescent↗
PubMed2000

High human plasma levels of organochlorine compounds in Greenland. Regional differences and lifestyle effects.

INTRODUCTION: The purpose of the study has been to analyse data, collected for surveillance purposes under the Human Health Programme of AMAP (Arctic Monitoring and Assessment Programme, in Greenland, regarding geographical differences and lifestyle versus pollution load from persistent organic pollutants (POP), and to make possible exposure assessments. It comprises a regional study from six districts and an ongoing study of pregnant women and infants. METHODOLOGY: Biostatistical analyses of data collected in Greenland from 61 men and 10 women from six different districts (1997-98) and from 110 mother-infant pairs in the Disko Bay area (1996-97) and 223 mother-infant pairs (1994-96). The data consisted of questionnaire answers and bloodsamples from men, women, and newborn infants (cord blood) analysed for fatty acids, selenium and 26 POPs including 14 PCB-congeners and four toxaphenes. RESULTS: Strong regional differences were found, related to different intakes of marine food with very high PCB-loads among men from the East coast. The various POP-plasma levels were mutually correlated and strong correlations were found between POP-plasma concentrations in mothers and new-born, R > 0.9, p < 0.0001. Among the pregnant women 95% surpassed the Canadian concern level for PCB, and 60% of the men from Scoresbysund surpassed the action level. The association between reported monthly food frequency and POPs was relatively weak, but the POPs were strongly correlated with plasma and erythrocyte n-3/n-6 fatty acid ratios as biomarkers of marine food intake. Multiple regression analysis showed highly significant positive correlation between smoking, and POP-plasma levels, after correction for age, alcohol intake, marine food, plasma lipids and n-3/n-6 fatty acid ratios. CONCLUSION: As the most important determinants of high POP-plasma levels in Greenlanders we propose: age, high plasma n-3 fatty acids (marine food), East coast region, and being a smoker.

Adult↗
PubMed1999

Functional interaction between GCN5 and polyamines: a new role for core histone acetylation.

Polyamines are organic polycations essential for a wide variety of cellular functions, including nuclear integrity and chromosome condensation. Here we present genetic evidence that depletion of cellular polyamines partially alleviates the defects in HO and SUC2 expression caused by inactivation of the GCN5 histone acetyltransferase. In addition, the combination of polyamine depletion and a sin(-) allele of the histone H4 gene leads to almost complete bypass of the transcriptional requirement for GCN5. In contrast, polyamine depletion does not alter the transcriptional requirements for the SWI/SNF chromatin remodeling complex nor does depletion lead to global defects in transcriptional regulation. In addition to these genetic studies, we show that polyamines facilitate oligomerization of nucleosomal arrays in vitro, and that polyamine-mediated condensation requires intact core histone N-terminal domains and is inhibited by histone hyperacetylation. Our studies suggest that polyamines are repressors of transcription in vivo, and that one role of histone hyperacetylation is to antagonize the ability of polyamines to stabilize highly condensed states of chromosomal fibers.

Acetylation↗
PubMed1999

The core histone N-terminal domains are required for multiple rounds of catalytic chromatin remodeling by the SWI/SNF and RSC complexes.

SWI/SNF and RSC are large, distinct multi-subunit complexes that use the energy of ATP hydrolysis to disrupt nucleosome structure, facilitating the binding of transcription factors or restriction enzymes to nucleosomes [Cote, J., Quinn, J., Workman, J. L., and Peterson, C. L. (1994) Science 265, 53-60 (1); Lorch, Y., Cairns, B. R., Zhang, M., and Kornberg, R. D. (1998) Cell 94, 29-34 (2)]. Here we have used a quantitative assay to measure the activities of these ATP-dependent chromatin remodeling complexes using nucleosomal arrays reconstituted with hypoacetylated, hyperacetylated, or partially trypsinized histones. This assay is based on measuring the kinetics of restriction enzyme digestion of a site located within the central nucleosome of a positioned 11-mer array [Logie, C., and Peterson, C. L. (1997) EMBO J. 16, 6772-6782 (3)]. We find that the DNA-stimulated ATPase activities of SWI/SNF and RSC are not altered by the absence of the histone N-termini. Furthermore, ATP-dependent nucleosome remodeling is also equivalent on all three substrate arrays under reaction conditions where the concentrations of nucleosomal array and either SWI/SNF or RSC are equivalent. However, SWI/SNF and RSC cannot catalytically remodel multiple nucleosomal arrays in the absence of the histone termini, and this catalytic activity of SWI/SNF is decreased by histone hyperacetylation. These results indicate that the histone termini are important for SWI/SNF and RSC function; and, furthermore, our data defines a step in the remodeling cycle where the core histone termini exert their influence. This step appears to be after remodeling, but prior to intermolecular transfer of the remodelers to new arrays.

Acetylation↗
PubMed1999

High serum coenzyme Q10, positively correlated with age, selenium and cholesterol, in Inuit of Greenland. A pilot study.

Greenlanders (Eskimos) have low prevalence of ischaemic heart disease, partly explained by a lower extent of atherosclerosis and a low n-6/n-3 ratio of polyunsaturated fatty acids. As atherosclerosis is also a result of oxidative stress, the total antioxidative readiness could have a substantial impact. From a health survey we chose the subpopulation from the most remote area, where the traditional Greenlandic diet with high intake of sea mammals and fish predominates. The mean (SD) of S-CoQ10 in males was 1.495 (0.529) nmol/ml and 1.421 (0.629) nmol/ml in females, significantly higher (p < 0.001) compared to a Danish population. In a linear multiple regression model the S-CoQ10 level is significantly positively associated with age and S-selenium in males, and S-total cholesterol in females. The high level of CoQ10 in Greenlanders probably reflects diet, since no bioaccumulation takes place, and it could probably be a substantial part of the antioxidative defense.

Adult↗
PubMed1999

Internal carotid artery patency following common carotid artery occlusion: management of the asymptomatic patient.

Rarely, internal carotid artery (ICA) patency is maintained above a common carotid artery occlusion (CCAO) through collateral flow of the ipsilateral external carotid artery or an aberrant ICA branch. Several small series report excellent results of ICA revascularization in relieving cerebral ischemic symptoms in these patients. The natural history of CCAO with patent ICA in the asymptomatic patient, however, is unknown. The Greenville Memorial Hospital Vascular Teaching Service registry and all carotid duplex scans done in the Greenville Hospital System vascular laboratory from January 1994 through December 1997 were reviewed. Data collection included chart review, phone interviews, and the review of angiograms and duplex scans. This study suggests that carotid duplex is more sensitive for detecting ICA flow after CCAO than routine contrast angiography. It also suggests that while rare in presentation, asymptomatic patients with CCAO and a patent ICA appear to have a benign neurologic course and can probably be observed without a high risk of stroke.

Carotid Artery, Common↗
PubMed1999

Concentration of organochlorines in human brain, liver, and adipose tissue autopsy samples from Greenland.

Organochlorines are persistent lipophilic compounds that accumulate in Inuit people living in circumpolar countries. Organochlorines accumulate as a result of the Inuits' large consumption of sea mammal fat; however, available data are limited to blood lipids, milk fat, and adipose tissue. We report results of organochlorine determination in liver, brain, omental fat, and subcutaneous abdominal fat samples collected from deceased Greenlanders between 1992 and 1994. Eleven chlorinated pesticides and 14 polychlorinated biphenyl congeners were measured in tissue lipid extracts by high-resolution gas chromatography with electron capture detection. Mean concentrations of polychlorinated biphenyls, 2, 2'-bis(4-chlorophenyl)-1,1-dichloroethylene, ss-hexachlorocyclohexane, hexachlorobenzene, mirex, trans-nonachlor, and oxychlordane in adipose tissue samples from Greenlanders were 3-34-fold higher than those measured using the same analytical method in samples from Canadians in Quebec City, Quebec. Brain lipids contained lower concentrations of all organochlorines than lipids extracted from other tissues. Organochlorine residue levels in lipid extracts from liver, omental fat, and subcutaneous abdominal fat samples were similar, with the exception of ss-hexachlorocyclohexane, which reached a greater concentration in liver lipids than in lipids from both adipose tissues (4-fold; p < 0. 05). Comparisons with available international data on adipose tissue levels reveal that the organochlorine body burden in the Inuit population of Greenland is presently among the highest resulting from environmental exposure.

Adipose Tissue↗