Prediction of neotropical tree and liana species richness from soil and climatic data

We present an analysis of local species richness in neotropical forests, based on a number of 0.1 ha samples of woody plants collected by the late Alwyn Gentry. For each of 69 forests, soils were analysed and climatic data were collated. Using transformed independent variables and interaction terms, multiple regression equations were developed that explained the greatest possible amount of variation in species richness, and the best equations were selected on the basis of regression diagnostics. The best models are presented for (a) all neotropical forests, (b) forests west of the Andes (transandean) and (c) east of the Andes (cisandean), and for various subsets based on elevation and annual rainfall. For the whole dataset, and for most subsets, annual rainfall and rainfall seasonality were the most important variables for explaining species richness. Soil variables were correlated with precipitation — drier forests have more nutrient-rich soils. After the inclusion of rainfall variables, available soil nutrient concentrations contributed little to explaining or accounting for additional variation in species numbers, indicating that tropical forest species richness is surprisingly independent of soil quality. The results are consistent with the hypothesis that plants in mature tropical forests may obtain nutrients through the process of direct cycling, in which mineral nutrients are extracted from litterfall before they enter the soil. The strong relationship between community species richness and rainfall patterns has implications for biodiversity conservation. Wet forests with an ample year-round moisture supply harbour the greatest number of woody plant species and should be a focus of conservation efforts.Died 3 August 1993.     

Common features of the yes and src gene products defined by peptide-specific antibodies.

Anti-peptide antibodies generated against a hydrophilic domain of pp60src comprising amino acid residues 498 through 512 were shown to be cross-reactive with the corresponding region in the yes transforming proteins encoded by Yamaguchi 73 and Esh sarcoma viruses. This cross-reactivity was demonstrated by immunoblot and immunoprecipitation analyses, and the identity of the proteins was verified by partial proteolytic mapping. By utilizing a combination of immunofluorescence and interference-reflection microscopy, these cross-reactive anti-peptide antibodies were shown to produce an immunofluorescence staining pattern in Yamaguchi 73 and Esh sarcoma virus-transformed chicken embryo fibroblasts remarkably similar to that pp60src in Rous sarcoma virus-infected chicken cells. Like the src gene products, the yes transformation-specific polyproteins were found to be concentrated within adhesion plaque structures and needle-like interdigitating cell-cell junctions. This analogous subcellular distribution suggests that these onc proteins are functionally related and may share common intracellular targets.     

Interaction of nucleolar phosphoprotein B23 with nucleic acids

The interaction of eukaryotic nucleolar phosphoprotein B23 with nucleic acids was examined by gel retardation and filter binding assays, by fluorescence techniques, and by circular dichroism. All studies utilized protein prepared under native conditions by a newly developed purification procedure. Electrophoretic gel mobility shift assays with phage M13 DNA suggested that protein B23 is a single-stranded nucleic acid binding protein. This was confirmed in competition binding assays with native or heat-denatured linearized plasmid pUC18 DNA where the protein showed a marked preference for the denatured form. In other competition assays, there was no apparent preference for single-stranded synthetic ribo- versus deoxyribonucleotides. Equilibrium binding with poly(riboethenoadenylic acid) indicated cooperative ligand binding with a protein binding site size of 11 nucleotides and an apparent binding constant (K omega) of 5 x 10(7) M-1 which includes an intrinsic binding constant (K) of 6.3 x 10(4) M-1 and a cooperativity factor (omega) of 800. In circular dichroism (CD) studies, protein B23, when combined with the single-stranded synthetic nucleic acids poly(rA) and poly(rC), effected a decrease in ellipticity and a shift of the positive peak at 260-270 nm toward higher wavelengths, indicating helix destabilizing activity. No CD changes were seen with double-stranded poly(dA.dT). The change in ellipticity of poly(rA) was sigmoidal upon addition of protein, confirming the cooperative behavior seen with fluorescence methods. These studies indicate that protein B23 binds cooperatively with high affinity for single-stranded nucleic acids and exhibits RNA helix destabilizing activity. These features may be related to its role in ribosome assembly.     

Specific coagulation factor adsorption to insoluble heparin

An insoluble derivative of heparin has been prepared by coupling to agarose beads, and its adsorption of certain blood clotting factors has been investigated. Factors IX^* XI, and heparin cofactor (He-Co)^** are selectively adsorbed, and can be recovered by elution of the heparin-agarose (H-Ag) with an NaCl gradient. Fibrinogen (FI) adsorption is negligible. The affinity for F IX appears to be lower than for F XI and He-Co. Thrombin, F XI, and He-Co seem to be equally tightly bound to the insoluble H.     

A functional interaction between the p75 neurotrophin receptor interacting factors, TRAF6 and NRIF

Neurotrophin signaling through the p75 receptor regulates apoptosis within the nervous system both during development and in response to injury. Whereas a number of p75 interacting factors have been identified, how these upstream factors function in a coordinated manner to mediate receptor signaling is still unclear. Here, we report a functional interaction between TRAF6 and the neurotrophin receptor interacting factor (NRIF), two proteins known to associate with the intracellular domain of the p75 neurotrophin receptor. The association between NRIF and TRAF6 was direct and occurred with both endogenous and ectopically expressed proteins. A KRAB repressor domain of NRIF and the carboxyl-terminal, receptor-binding region of TRAF6 were required for the interaction. Co-expression of TRAF6 increased the levels of NRIF protein and induced its nuclear translocation. Reciprocally, NRIF enhanced TRAF6-mediated activation of the c-Jun NH2-terminal kinase (JNK) by 3-fold, while only modestly increasing the stimulation of NF-kappaB. The expression of both NRIF and TRAF6 was required for reconstituting p75 activation of JNK in HEK293 cells, whereas NRIF mutants lacking the TRAF6 interaction domain were unable to substitute for the full-length protein in facilitating activation of the kinase. These results suggest that NRIF and TRAF6 functionally interact to facilitate neurotrophin signaling through the p75 receptor.     

MSH2 missense mutations alter cisplatin cytotoxicity and promote cisplatin-induced genome instability

Defects in the mismatch repair protein MSH2 cause tolerance to DNA damage. We report how cancer-derived and polymorphic MSH2 missense mutations affect cisplatin cytotoxicity. The chemotolerance phenotype was compared with the mutator phenotype in a yeast model system. MSH2 missense mutations display a strikingly different effect on cell death and genome instability. A mutator phenotype does not predict chemotolerance or vice versa. MSH2 mutations that were identified in tumors (Y109C) or as genetic variations (L402F) promote tolerance to cisplatin, but leave the initial mutation rate of cells unaltered. A secondary increase in the mutation rate is observed after cisplatin exposure in these strains. The mutation spectrum of cisplatin-resistant mutators identifies persistent cisplatin adduction as the cause for this acquired genome instability. Our results demonstrate that MSH2 missense mutations that were identified in tumors or as polymorphic variations can cause increased cisplatin tolerance independent of an initial mutator phenotype. Cisplatin exposure promotes drug-induced genome instability. From a mechanistical standpoint, these data demonstrate functional separation between MSH2-dependent cisplatin cytotoxicity and repair. From a clinical standpoint, these data provide valuable information on the consequences of point mutations for the success of chemotherapy and the risk for secondary carcinogenesis.     

Cellular localization of the Escherichia coli SpoT protein.

The SpoT protein of Escherichia coli serves as a source of degradation as well as an apparent source of synthesis of (p)ppGpp. Since the subcellular localization of SpoT might be a clue to its function, we have used SpoT-specific antisera to analyze cell extracts fractionated on sucrose gradients. We find that the SpoT protein is not bound to ribosomes or to either inner or outer membrane fractions. Although the SpoT protein is found in large aggregates, its localization is probably cytosolic.     

Nitration of polypeptides using ethanol in reaction buffers minimizes crosslinking.

High concentrations of ethanol are effective in reducing the intermolecular cross-linking which predominates when the reactions of tetranitromethane with the carboxypeptidase inhibitor from potatoes, a chymotrypsin inhibitor also from potatoes, and insulin are carried out under standard conditions. Including reagents such as ethanol may be of general utility in the preparation of monomeric nitro-derivatives of hydrophobic, low-molecular-weight proteins.