Protein interaction mapping in C. elegans using proteins involved in vulval development

Protein interaction mapping using large-scale two-hybrid analysis has been proposed as a way to functionally annotate large numbers of uncharacterized proteins predicted by complete genome sequences. This approach was examined in Caenorhabditis elegans, starting with 27 proteins involved in vulval development. The resulting map reveals both known and new potential interactions and provides a functional annotation for approximately 100 uncharacterized gene products. A protein interaction mapping project is now feasible for C. elegans on a genome-wide scale and should contribute to the understanding of molecular mechanisms in this organism and in human diseases.     

Molecular cloning and functional expression of glutamate receptor subunit genes

Three closely related genes, GluR1, GluR2, and GluR3, encode receptor subunits for the excitatory neurotransmitter glutamate. The proteins encoded by the individual genes form homomeric ion channels in Xenopus oocytes that are sensitive to glutamatergic agonists such as kainate and quisqualate but not to N-methyl-D-aspartate, indicating that binding sites for kainate and quisqualate exist on single receptor polypeptides. In addition, kainate-evoked conductances are potentiated in oocytes expressing two or more of the cloned receptor subunits. Electrophysiological responses obtained with certain subunit combinations show agonist profiles and current-voltage relations that are similar to those obtained in vivo. Finally, in situ hybridization histochemistry reveals that these genes are transcribed in shared neuroanatomical loci. Thus, as with gamma-aminobutyric acid, glycine, and nicotinic acetylcholine receptors, native kainate-quisqualate-sensitive glutamate receptors form a family of heteromeric proteins.     

Substrate quality and the temperature sensitivity of soil organic matter decomposition

Determining the relative temperature sensitivities of the decomposition of the different soil organic matter (SOM) pools is critical for predicting the long-term impacts of climate change on soil carbon (C) storage. Although kinetic theory suggests that the temperature sensitivity of SOM decomposition should increase with substrate recalcitrance, there remains little empirical evidence to support this hypothesis. In the study presented here, sub-samples from a single bulk soil sample were frozen and sequentially defrosted to produce samples of the same soil that had been incubated for different lengths of time, up to a maximum of 124 days. These samples were then placed into an incubation system which allowed CO₂ production to be monitored constantly and the response of soil respiration to short-term temperature manipulations to be investigated. The temperature sensitivity of soil CO₂ production increased significantly with incubation time suggesting that, as the most labile SOM pool was depleted the temperature sensitivity of SOM decomposition increased. This study is therefore one of the first to provide empirical support for kinetic theory. Further, using a modelling approach, we demonstrate that it is the temperature sensitivity of the decomposition of the more recalcitrant SOM pools that will determine long-term soil-C losses. Therefore, the magnitude of the positive feedback to global warming may have been underestimated in previous modelling studies.