Annually resolved ice core records of tropical climate variability over the past ~1800 years.

Ice cores from low latitudes can provide a wealth of unique information about past climate in the tropics, but they are difficult to recover and few exist. Here, we report annually resolved ice core records from the Quelccaya ice cap (5670 meters above sea level) in Peru that extend back ~1800 years and provide a high-resolution record of climate variability there. Oxygen isotopic ratios (δ(18)O) are linked to sea surface temperatures in the tropical eastern Pacific, whereas concentrations of ammonium and nitrate document the dominant role played by the migration of the Intertropical Convergence Zone in the region of the tropical Andes. Quelccaya continues to retreat and thin. Radiocarbon dates on wetland plants exposed along its retreating margins indicate that it has not been smaller for at least six millennia.

Identification of signals required for the insertion of heterologous genome segments into the reovirus genome.

In cells simultaneously infected with any two of the three reovirus serotypes ST1, ST2, and ST3, up to 15% of the yields are intertypic reassortants that contain all possible combinations of parental genome segments. We have now found that not all genome segments in reassortants are wild type. In reassortants that possess more ST1 than ST3 genome segments, all ST1 genome segments appear to be wild type, but the incoming ST3 genome segments possess mutations that make them more similar to the ST1 genome segments that they replace. In reassortants resulting from crosses of the more distantly related ST3 and ST2 viruses that possess a majority of ST3 genome segments, all incoming ST2 genome segments are wild type, but the ST3 S4 genome segment possesses two mutations, G74 to A and G624 to A, that function as acceptance signals. Recognition of these signals has far-reaching implications for the construction of reoviruses with novel properties and functions.

Late glacial stage and holocene tropical ice core records from huascaran, peru.

Two ice cores from the col of Huascarán in the north-central Andes of Peru contain a paleoclimatic history extending well into the Wisconsinan (Würm) Glacial Stage and include evidence of the Younger Dryas cool phase. Glacial stage conditions at high elevations in the tropics appear to have been as much as 8 degrees to 12 degrees C cooler than today, the atmosphere contained about 200 times as much dust, and the Amazon Basin forest cover may have been much less extensive. Differences in both the oxygen isotope ratio zeta(18)O (8 per mil) and the deuterium excess (4.5 per mil) from the Late Glacial Stage to the Holocene are comparable with polar ice core records. These data imply that the tropical Atlantic was possibly 5 degrees to 6 degrees C cooler during the Late Glacial Stage, that the climate was warmest from 8400 to 5200 years before present, and that it cooled gradually, culminating with the Little Ice Age (200 to 500 years before present). A strong warming has dominated the last two centuries.

The influence of the base-paired flanking region on structure and function of the ferritin mRNA iron regulatory element.

Ferritin and transferrin receptors are co-ordinately regulated by the same RNA-protein interaction: the conserved iron regulatory element (IRE) in mRNA and the IRE-binding protein (IRE-BP/IRP/FRP/P-90). The 28 nucleotide IRE in ferritin mRNA is a single copy, with base-paired flanking regions (FL), located near the 5' cap. In the transferrin receptor mRNA, the IRE is located in the 3' untranslated region, as five variable copies and lacking predicted base-paired flanking regions; an alternate predicted structure without IREs has similar stability. When iron is scarce, ferritin mRNA does not form polyribosomes whereas the transferrin receptor mRNA is translated; when iron is abundant, ferritin mRNA forms polyribosomes and the transferrin receptor mRNA is degraded. To investigate structures which contribute to differences in the regulation of the two mRNAs, the effect of mutation of the ferritin FL was studied. Changes in structure (changes in reactivity with RNase V1 and RNase S1. Fe-bleomycin) and changes in function (translation in rabbit reticulocyte extracts) were compared for mutant and wild-type FL sequences in ferritin mRNA. The disruption of a triplet of base-pairs in the FL had diminished regulation; a second mutation to restore the triplet base-pairs conferred wild-type translational regulation. Conformation of the mutant RNA-IRE-BP complex was also different. We show that the triplet of base-pairs is conserved; the triplet is also the location of IRE-BP-dependent conformational changes in the FL structure previously observed. Increasing FL base-pairs had no effect on function. Structural changes associated with altered function included bleomycin sites in the IRE, suggesting an alternate conformation of the hairpin, and different base-stacking (V1 sensitivity) in the FL. The function of the FL, which is altered by mutation of phylogenetically conserved triplet base-pairs, may be enhancement of formation of a particular IRE stem-loop-protein interaction.

The iron regulatory region of ferritin mRNA is also a positive control element for iron-independent translation.

The iron regulatory element (IRE) in the 5'-untranslated region of ferritin mRNA interacts with a specific regulator protein (P-90, IRE-BP, or FRP) to block translation. High cellular iron changes the IRE/P-90 interaction to relax the translational block and allow polyribosome formation. We now show that the IRE and base-paired flanking regions also enhance translation in the absence of P-90, explaining the high translational efficiency of deregulated ferritin mRNA observed previously. The effect of the IRE on translational efficiency was examined by comparing four sets of mRNAs: (1) +/- IRE in animal (frog) ferritin, regulated translationally by iron in vivo; (2) +/- animal IRE fused with plant (soybean) ferritin, regulated transcriptionally by iron in vivo; (3) repositioned IRE in animal ferritin; (4) mutated IRE in animal ferritin with G16A substitution, which decreases P-90 binding (negative control). The IRE region increased translational efficiency of both the animal ferritin and the heterologous IRE/soybean ferritin fusion mRNAs; the effect was observed in cell-free translation systems from either plants (wheat germ) or animals (rabbit reticulocyte). Repositioning the IRE further from the 5' cap eliminated positive control of translation. The single base mutation had no effect, indicating that positive and negative translational control involves different sections of the IRE region. Thus, the IRE region in ferritin mRNA encodes both positive translational control and, when combined with the regulator protein P-90, negative translational control.

Ferritin mRNA probed, near the iron regulatory region, with protein and chemical (1,10-phenanthroline-Cu) nucleases. A possible role for base-paired flanking regions.

Iron stimulates ferritin synthesis in whole cells and animals, by increasing the entry of ferritin mRNA into polyribosomes. Dissection of the regulation at the molecular level has identified a 28-nucleotide, conserved, regulatory sequence (IRE = iron regulatory element) in the 5' non-coding region of ferritin mRNAs, plus trans-acting factor(s), one of which is a 90-kDa protein. The site of iron action is not entirely characterized but may involve heme; sequences in the 3' non-coding region of ferritin mRNA can modulate regulation. Ferritin mRNA is the first eukaryotic mRNA for which a conserved regulatory sequence and regulator protein have been identified. The same RNA-protein motif is used, through iron-dependent degradation of transferrin receptor mRNA, to decrease synthesis of the receptor and cellular iron uptake. The regulatory structure of the transferrin receptor mRNA is composed, in part, of five copies of the IRE in the 3' non-coding region. IRE structure, probed by cleavage with RNases T1, V1, 1,10-phenanthroline-Cu or modification with dimethyl sulfate, is a hairpin loop with conformational variations dependent on magnesium; a base-paired region flanking the IRE is also structurally sensitive to magnesium. Similar results were obtained with a synthetic 55-mer containing the IRE and with a full-length in vitro transcript with a G----A substitution in the loop.(ABSTRACT TRUNCATED AT 250 WORDS)

[Isolation of polynectin and its comparison with fibronectin].

A gelatin-binding protein polynectin (PN) was isolated from porcine plasma by affinity chromatography in sequence of columns of Sepharose 4B coupled with gelatin, arginine and heparin and then by gel filtration through Sephadex G-200. This protein was bound strongly to arginine column and thus could be separated from fibronectin (FN). PN is similar to FN with respect to bind characteristics to affinity gels and with a molecular weight of about 450,000 in PAGE, PN being composed of several small subunits of the same molecular weight linked by disulfide bonds. Results of immunodiffusion and ELISA shows that: no immune reaction was observed between anti-PN antibody and FN and PN existed only in porcine plasma. but not in other animal (bovine, goat, mouse and rat) or human plasma. PN exhibited no cell-attachment-promoting effects on CHO cell. These indicate that PN was a glycoprotein different from FN. Moreover, a high titer (1:32) rabbit antiserum against porcine plasma FN (free from PN) was prepared from immunized rabbit, from which affinity purified anti-FN antibody was obtained through FN-Sepharose 4B column.

[Isolation and characterization of fibronectin from porcine plasma].

Electrophoretic purified fibronectin (FN) was isolated with high concentration (0.5 mg/ml plasma) from porcine plasma by affinity chromatography with gelatin-sepharose 4B and heparin-sepharose 4B. The isolated FN shows one single band (MW 450,000) both in agarose gel and PAGE, and two similar bands (MW 230,000) in the presence of 1% beta-mercaptoethanol in SDS-PAGE. The FN isolated remains its immunological and biological properties, being reactable with antibodies against human and bovine FN with strong promotion effect on CHO cell adhesion in serum free medium. These data indicate that porcine plasma is a good resource for isolation of FN.