Binding of SH2 domains of phospholipase C gamma 1, GAP, and Src to activated growth factor receptors

Phospholipase C gamma 1 (PLC gamma 1) and p21ras guanosine triphosphatase (GTPase) activating protein (GAP) bind to and are phosphorylated by activated growth factor receptors. Both PLC gamma 1 and GAP contain two adjacent copies of the noncatalytic Src homology 2 (SH2) domain. The SH2 domains of PLC gamma 1 synthesized individually in bacteria formed high affinity complexes with the epidermal growth factor (EGF)- or platelet derived growth factor (PDGF)-receptors in cell lysates, and bound synergistically to activated receptors when expressed together as one bacterial protein. In vitro complex formation was dependent on prior growth factor stimulation and was competed by intracellular PLC gamma 1. Similar results were obtained for binding of GAP SH2 domains to the PDGF-receptor. The isolated SH2 domains of other signaling proteins, such as p60src and Crk, also bound activated PDGF-receptors in vitro. SH2 domains, therefore, provide a common mechanism by which enzymatically diverse regulatory proteins can physically associate with the same activated receptors and thereby couple growth factor stimulation to intracellular signal transduction pathways.     

Phosphorylation networks regulating JNK activity in diverse genetic backgrounds

Cellular signaling networks have evolved to enable swift and accurate responses, even in the face of genetic or environmental perturbation. Thus, genetic screens may not identify all the genes that regulate different biological processes. Moreover, although classical screening approaches have succeeded in providing parts lists of the essential components of signaling networks, they typically do not provide much insight into the hierarchical and functional relations that exist among these components. We describe a high-throughput screen in which we used RNA interference to systematically inhibit two genes simultaneously in 17,724 combinations to identify regulators of Drosophila JUN NH(2)-terminal kinase (JNK). Using both genetic and phosphoproteomics data, we then implemented an integrative network algorithm to construct a JNK phosphorylation network, which provides structural and mechanistic insights into the systems architecture of JNK signaling.     

Large-scale,multi-year,phenology modelling of forest insects in Pinus radiata plantations

Journal of Pest Science - Ecologically and economically harmful wood borers and bark beetles, which have the capacity to expand geographically through the international log trade, require...     

The effects of crowding on mackerel (Scomber scombrus L.) — Physical condition and mortality

Series of trials in which mackerel (Scomber scombrus L.) were confined in keepnets at different stocking densities are described. From simple confinement trials it was found that 50% of the fish died after 48 h at a stocking density of 30 fish m⁻³, equivalent to 6.5 kg m⁻³. Trials in which fish were held at stocking densities, and for a duration, comparable to those experienced in a “dried up” purse seine prior to “slipping”, showed that up to 90% of “slipped” fish died within 48 h of release. The primary cause of death was probably physical damage, particularly skin loss, caused by abrasion, although there is some evidence that mackerel have a healing process which can accommodate minor skin abrasions. A tagging trial showed a small but significant increase in mortality due to the tagging procedure.     

A comparison of the effects of two different doses of ketamine used for co-induction of anaesthesia with a target-controlled infusion of propofol in dogs

ObjectiveTo assess the cardiorespiratory and hypnotic-sparing effects of ketamine co-induction with target-controlled infusion of propofol in dogs.Study designProspective, randomized, blinded clinical study.AnimalsNinety healthy dogs (ASA grades I/II). Mean body mass 30.5 ± SD 8.6 kg and mean age 4.2 ± 2.6 years.MethodsAll dogs received pre-anaesthetic medication with acepromazine (0.03 mg kg^?1) and morphine (0.2 mg kg^?1) administered intramuscularly 30 minutes prior to induction of anaesthesia. Heart rate and respiratory rate were recorded prior to pre-medication. Animals were allocated into three different groups: Group 1 (control) received 0.9% NaCl, group 2, 0.25 mg kg^?1 ketamine and group 3, 0.5 mg kg^?1 ketamine, intravenously 1 minute prior to induction of anaesthesia, which was accomplished using a propofol target-controlled infusion system. The target propofol concentration was gradually increased until endotracheal intubation was possible and the target concentration at intubation was recorded. Heart rate, respiratory rate and noninvasive blood pressure were recorded immediately prior to induction, at successful intubation and at 3 and 5 minutes post-intubation. The quality of induction was graded according to the amount of muscle twitching and paddling observed. Data were analysed using a combination of chi-squared tests, Fisher's exact tests, Kruskal–Wallis, and anova with significance assumed at p< 0.05.ResultsThere were no significant differences between groups in the blood propofol targets required to achieve endotracheal intubation, nor with respect to heart rate, noninvasive blood pressure or quality of induction. Compared with the other groups, the incidence of post-induction apnoea was significantly higher in group 3, but despite this dogs in this group had higher respiratory rates overall.Conclusions and clinical relevanceUnder the conditions of this study, ketamine does not seem to be a useful agent for co-induction of anaesthesia with propofol in dogs.     

X-ray Diffraction Studies of Echinoderm Plates

X-ray diffraction studies confirm that, with few exceptions, each skeletal element of echtinoderms is a single crystal of magnesium-rich calcite and that a relation exists between the shape of the element and the crystallographic a- and c-axes. The exceptions incluide the teeth of echinoids, and the calcareous ring as well as the anal teeth of holothurians. The tubercles of an echinoid plate begin their growth as parts of the single crystal of the plate; under the mechanical action of the spines that are attached to them, they become partly polycrystalline, as shown by scanning electron microscopy and by x-ray powder diffraction. The interface between inorganic crystalline and organic amorphous matter in the skeletal element appears to be the first example reported in nature of a periodic mninimal suirface.     

Combinatorial ShcA docking interactions support diversity in tissue morphogenesis

Changes in protein-protein interactions may allow polypeptides to perform unexpected regulatory functions. Mammalian ShcA docking proteins have amino-terminal phosphotyrosine (pTyr) binding (PTB) and carboxyl-terminal Src homology 2 (SH2) domains, which recognize specific pTyr sites on activated receptors, and a central region with two phosphorylated tyrosine-X-asparagine (pYXN) motifs (where X represents any amino acid) that each bind the growth factor receptor-bound protein 2 (Grb2) adaptor. Phylogenetic analysis indicates that ShcA may signal through both pYXN-dependent and -independent pathways. We show that, in mice, cardiomyocyte-expressed ShcA directs mid-gestational heart development by a PTB-dependent mechanism that does not require the pYXN motifs. In contrast, the pYXN motifs are required with PTB and SH2 domains in the same ShcA molecule for the formation of muscle spindles, skeletal muscle sensory organs that regulate motor behavior. Thus, combinatorial differences in ShcA docking interactions may yield multiple signaling mechanisms to support diversity in tissue morphogenesis.