Yeast DNA polymerase epsilon participates in leading-strand DNA replication

Multiple DNA polymerases participate in replicating the leading and lagging strands of the eukaryotic nuclear genome. Although 50 years have passed since the first DNA polymerase was discovered, the identity of the major polymerase used for leading-strand replication is uncertain. We constructed a derivative of yeast DNA polymerase epsilon that retains high replication activity but has strongly reduced replication fidelity, particularly for thymine-deoxythymidine 5'-monophosphate (T-dTMP) but not adenine-deoxyadenosine 5'-monophosphate (A-dAMP) mismatches. Yeast strains with this DNA polymerase epsilon allele have elevated rates of T to A substitution mutations. The position and rate of these substitutions depend on the orientation of the mutational reporter and its location relative to origins of DNA replication and reveal a pattern indicating that DNA polymerase epsilon participates in leading-strand DNA replication.     

Experimental verification of a negative index of refraction

We present experimental scattering data at microwave frequencies on a structured metamaterial that exhibits a frequency band where the effective index of refraction (n) is negative. The material consists of a two-dimensional array of repeated unit cells of copper strips and split ring resonators on interlocking strips of standard circuit board material. By measuring the scattering angle of the transmitted beam through a prism fabricated from this material, we determine the effective n, appropriate to Snell's law. These experiments directly confirm the predictions of Maxwell's equations that n is given by the negative square root of epsilon.mu for the frequencies where both the permittivity (epsilon) and the permeability (mu) are negative. Configurations of geometrical optical designs are now possible that could not be realized by positive index materials.     

Autonomous developmental control of human embryonic globin gene switching in transgenic mice

The mechanisms by which expression of the beta-like globin genes are developmentally regulated are under intense investigation. The temporal control of human embryonic (epsilon) globin expression was analyzed. A 3.7-kilobase (kb) fragment that contained the entire human epsilon-globin gene was linked to a 2.5-kb cassette of the locus control region (LCR), and the developmental time of expression of this construct was studied in transgenic mice. The human epsilon-globin transgene was expressed in yolk sac-derived primitive erythroid cells, but not in fetal liver or bone marrow-derived definitive erythroid cells. The absence of epsilon gene expression in definitive erythroid cells suggests that the developmental regulation of the epsilon-globin gene depends only on the presence of the LCR and the epsilon-globin gene itself (that is, an autonomous negative control mechanism). The autonomy of epsilon-globin gene developmental control distinguishes it from the competitive mechanism of regulation of gamma and beta-globin genes, and therefore, suggests that at least two distinct mechanisms function in human hemoglobin switching.     

Experimental Evidence for a New Iron Phase and Implications for Earth's Core

Iron is known to occur in four different crystal structural forms. One of these, the densest form (epsilon phase, hexagonal close-packed) is considered to have formed Earth's core. Theoretical arguments based on available high-temperature and high-pressure iron data indicate the possibility of a fifth less dense iron phase forming the core. Study of iron phase transition conducted between pressures of 20 to 100 gigapascals and 1000 to 2200 Kelvin provides an experimental confirmation of the existence of this new phase. Thee epsilon iron phase transforms to this lower density phase before melting. The new phase may form a large part of Earth's core.     

Rhenium-osmium and samarium-neodymium isotopic systematics of the stillwater complex

Isotopic data for the Stillwater Complex, Montana, which formed about 2700 Ma (million years ago), were obtained to evaluate the role of magma mixing in the formation of strategic platinum-group element (PGE) ore deposits. Neodymium and osmium isotopic data indicate that the intrusion formed from at least two geochemically distinct magmas. Ultramafic affinity (U-type) magmas had initial epsilon(Nd) of -0.8 to -3.2 and a chondritic initial (187)Os/(186)Os ratio of approximately 0.88, whereas anorthositic affinity (A-type) magmas had epsilon(Nd) of -0.7 to +1.7 and an initial (187)Os/(186)Os ratio of approximately -1.13. These data suggest that U-type magmas were derived from a lithospheric mantle source containing recycled crustal materials whereas A-type magmas originated either by crustal contamination of basaltic magmas or by partial melting of basalt in the lower crust. The Nd and Os isotopic data also suggest that Os, and probably the other PGEs in ore horizons such as the J-M Reef, was derived from A-type magmas. The Nd and Os isotopic heterogeneity observed in rocks below the J-M Reef also suggests that A-type magmas were injected into the Stillwater U-type magma chamber at several stages during the development of the Ultramafic series.     

Synchrotron X-ray Study of Iron at High Pressure and Temperature

X-ray synchrotron experiments with in situ laser heating of iron in a diamond-anvil cell show that the high-pressure epsilon phase, a hexagonal close-packed (hcp) structure, transforms to another phase (possibly a polytype double-layer hcp) at a pressure of about 38 gigapascals and at temperatures between 1200 and 1500 kelvin. This information has implications for the phase relations of iron in Earth's core.     

Hemoglobin Portland 1: a new human hemoglobin unique in structure

A new hemoglobin (Hb), Portland 1, has been found in a newborn infant having multiple congenital anomalies and complex autosomal chromosomal mosaicism. The new hemoglobin has a unique tetrameric structure (molecular weight, 66,000) composed of two pairs of different types of chains, neither of which is alpha, gamma(2)x(2). The x-chain of Hb Portland 1 may be a new type of hemoglobin chain, but the available evidence suggests that it may be identical with the epsilon chain. We suggest that Hb Portland 1 is an embryonic hemoglobin that persisted until after birth in relatively large amounts in this patient.     

Mechanisms for regulating expression of membrane isoforms of Fc gamma RIII (CD16)

Granulocyte and natural killer (NK) cell Fc receptors for immunoglobulin G (CD16) differ in only a few amino acids, yet have phosphatidylinositol glycan (PIG) or polypeptide membrane anchors, respectively. Mutagenesis shows that anchoring is regulated by a serine residue near the PIG anchor attachment site in the extracellular domain. The NK cell isoform was not expressed on the surface of COS cells unless cotransfected with a subunit that was expressed in NK cells and that was identical to the gamma subunit of the high affinity IgE Fc receptor (Fc epsilon RI). However, the CD16 sequence and not expression of the gamma subunit is dominant in regulating PIG reanchoring.     

Mechanism of inactivation of enzyme proteins by ultraviolet light

Some quantum yields for the destruction of amino acids have been determined. The inactivation of the enzymes hymotrypsin, lysozyme, ribonuclease, and trypsin by ultraviolet light can be accounted for quantitatively by summing the products of (i) the probability that light is absorbed by a given amino acid residue, epsilon(4), and (ii) the probability that absorbed light induces a chemical change, with a quantum efficiency phi(4), in the residue. The principal residues involved are cystyl and tryptophanyl. Peptide bond rupture is not important. Analysis of inactivated enzymes verifies the assumption of the existence of several inactivation mechanisms.     

Cross-link in fibrin polymerized by factor 13: epsilon-(gamma-glutamyl)lysine

(epsilon)-((gamma)-Glutamyl)lysine has been isolated from enzymatic hydrolyzates of cross-linked human fibrin. This compound was not detected in "non-cross-linked" fibrin prepared with ethylenediaminetetraacetic acid, which inhibits factor XIII; intermediate amounts were observed when the fibrin was prepared with glycine ethyl ester, which inhibits factor XIII competitively. These and ancillary experiments furnish conclusive evidence that epsilon-(gamma-glutamyl)lysine cross-links form in human fibrin during polymerization catalyzed by factor XIII.     

Asymmetrical allocation of NMDA receptor epsilon2 subunits in hippocampal circuitry

Despite its implications for higher order functions of the brain, little is currently known about the molecular basis of left-right asymmetry of the brain. Here we report that synaptic distribution of N-methyl-D-aspartate (NMDA) receptor GluRepsilon2 (NR2B) subunits in the adult mouse hippocampus is asymmetrical between the left and right and between the apical and basal dendrites of single neurons. These asymmetrical allocations of epsilon2 subunits differentiate the properties of NMDA receptors and synaptic plasticity between the left and right hippocampus. These results provide a molecular basis for the structural and functional asymmetry of the mature brain.     

The origin of aluminum flocs in polluted streams

About 240,000 square kilometers of Earth's surface is disrupted by mining, which creates watersheds that are polluted by acidity, aluminum, and heavy metals. Mixing of acidic effluent from old mines and acidic soils into waters with a higher pH causes precipitation of amorphous aluminum oxyhydroxide flocs that move in streams as suspended solids and transport adsorbed contaminants. On the basis of samples from nine streams, we show that these flocs probably form from aggregation of the epsilon -Keggin polyoxocation AlO4Al12(OH)24(H2O)12(7+)(aq) (Al13), because all of the flocs contain distinct Al(O)4 centers similar to that of the Al13 nanocluster.     

Atmospheric oxygen: isotopic composition and solubility fractionation

Atmospheric oxygen has been found to be enriched in oxygen-18 by 23.5 +/- 0.3 per mil relative to average ocean water (SMOW). Oxygen dissolved in seawater is further enriched in oxygen-18 by 0.85 per mil at 0 degrees C. The temperature dependence of the solubility enrichment is given by epsilon (per mil) = 0.85- 0.010 t ( degrees C). This result is in good agreement with earlier measurements of the solubility effect in distilled water.     

Heterogeneity of 5S RNA in fungal ribosomes

Neurospora crassa has at least seven types of 5S RNA genes (alpha, beta, gamma, epsilon, delta, zeta, and eta) with different coding regions. A high resolution gel electrophoresis system was developed to separate minor 5S RNA's from the major 5S RNA (alpha). A study of several Neurospora crassa strains, four other species in the genus Neurospora, members of two closely related genera, and three distantly related genera demonstrated that 5S RNA heterogeneity is common among fungi. In addition, different 5S RNA's are present in Neurospora ribosomes. The finding that fungal ribosomes are structurally heterogeneous suggests that ribosomes may be functionally heterogeneous as well.     

HEL cells: a new human erythroleukemia cell line with spontaneous and induced globin expression

A new human erythroleukemia cell line has been established. This line, designated HEL, is capable of spontaneous and induced globin synthesis, producing mainly G gamma and A gamma chains. Embryonic chains (epsilon, zeta) and alpha chains are detectable in very small amounts; beta chains are undetectable. This line provides a new model system for studying aspects of erythroid cell differentiation and differential globin gene expression.     

Neodymium-142 evidence for Hadean mafic crust

Neodymium-142 data for rocks from the Nuvvuagittuq greenstone belt in northern Quebec, Canada, show that some rock types have lower 142Nd/144Nd ratios than the terrestrial standard (epsilon142Nd = -0.07 to -0.15). Within a mafic amphibolite unit, 142Nd/144Nd ratios correlate positively with Sm/Nd ratios and produce a 146Sm-142Nd isochron with an age of 4280(-81)(+53) million years. These rocks thus sample incompatible-element-enriched material formed shortly after Earth formation and may represent the oldest preserved crustal section on Earth.     

Heterogeneous Hadean hafnium: evidence of continental crust at 4.4 to 4.5 ga

The long-favored paradigm for the development of continental crust is one of progressive growth beginning at approximately 4 billion years ago (Ga). To test this hypothesis, we measured initial 176Hf/177Hf values of 4.01- to 4.37-Ga detrital zircons from Jack Hills, Western Australia. epsilonHf (deviations of 176Hf/177Hf from bulk Earth in parts per 10(4)) values show large positive and negative deviations from those of the bulk Earth. Negative values indicate the development of a Lu/Hf reservoir that is consistent with the formation of continental crust (Lu/Hf approximately 0.01), perhaps as early as 4.5 Ga. Positive epsilon(Hf) deviations require early and likely widespread depletion of the upper mantle. These results support the view that continental crust had formed by 4.4 to 4.5 Ga and was rapidly recycled into the mantle.     

Origin of the sudbury complex by meteoritic impact: neodymium isotopic evidence

Samarium-neodymium isotopic data on whole rocks and minerals of the Sudbury Complex in Canada gave an igneous crystallization age of 1840 +/- 21 x 10(6) years. The initial epsilon neodymium values for 15 whole rocks are similar to those for average upper continental crust, falling on the crustal trend of neodymium isotopic evolution as defined by shales. The rare earth element concentration patterns of Sudbury rocks are also similar to upper crustal averages. These data suggest that the Sudbury Complex formed from melts generated in the upper crust and are consistent with a meteoritic impact.     

Isotope-edited NMR of cyclosporin A bound to cyclophilin: evidence for a trans 9,10 amide bond

The binding of a 13C-labeled cyclosporin A (CsA) analog to cyclophilin (peptidyl prolyl isomerase) was examined by means of isotope-edited nuclear magnetic resonance (NMR) techniques. A trans 9,10 peptide bond was adopted when CsA was bound to cyclophilin, in contrast to the cis 9,10 peptide bond found in the crystalline and solution conformations of CsA. Furthermore, nuclear Overhauser effects (NOEs) were observed between the zeta 3 and epsilon 3 protons of the methylleucine (MeLeu) residue at position 9 of CsA and tryptophan121 (Trp121) and phenylalanine (Phe) protons of cyclophilin, suggesting that the MeLeu9 residue of CsA interacts with cyclophilin. These results illustrate the power of isotope-edited NMR techniques for rapidly providing useful information about the conformations and active site environment of inhibitors bound to their target enzymes.