Role of the Sec61 translocon in EGF receptor trafficking to the nucleus and gene expression

The epidermal growth factor (EGF)-dependent trafficking of the intact EGF receptor to the nucleus and its requirement for growth factor induction of cyclin D and other genes has been reported. Unresolved is the mechanism by which this or other transmembrane proteins are excised from a lipid bilayer before nuclear translocalization. We report that, after the addition of EGF, the cell surface EGF receptor is trafficked to the endoplasmic reticulum (ER) where it associates with Sec61beta, a component of the Sec61 translocon, and is retrotranslocated from the ER to the cytoplasm. Abrogation of Sec61beta expression prevents EGF-dependent localization of EGF receptors to the nucleus and expression of cyclin D. This indicates that EGF receptors are trafficked from the ER to the nucleus by a novel pathway that involves the Sec61 translocon.     

High-level bacterial expression and purification of apicomplexan micronemal proteins for structural studies

The cysteine-rich N-terminal domain of the micronemal adhesive protein MIC1 (MIC1-NT) from Toxoplasma gondii was cloned, expressed in Escherichia coli and purified. MIC1-NT is amenable to structural studies as shown by preliminary NMR and X-ray analysis. Positive results with two further micronemal proteins indicate that our strategy has wider application.     

Segmental Aneuploidy and the Genetic Gross Structure of the Drosophila Genome

By combining elements of two Y-autosome translocations with displaced autosomal breakpoints, it is possible to produce zygotes heterozygous for a deficiency for the region between the breakpoints, and also, as a complementary product, zygotes carrying a duplication for precisely the same region. A set of Y-autosome translocations with appropriately positioned breakpoints, therefore, can in principle be used to generate a non-overlapping set of deficiencies and duplications for the entire autosomal complement.-Using this method, we have succeeded in examining segmental aneuploids for 85% of chromosomes 2 and 3 in order to assess the effects of aneuploidy and to determine the number and location of dosage-sensitive loci in the Drosophila genome (Figure 5). Combining our data with previously reported results on the synthesis of Drosophila aneuploids (see Lindsley and Grell 1968), the following generalities emerge.-1. The X chromosome contains no triplo-lethal loci, few or no haplo-lethal loci, at least seven Minute loci, one hyperploid-sensitive locus, and one locus that is both triplo-abnormal and haplo-abnormal. 2. Chromosome 2 contains no triplo-lethal loci, few or no haplo-lethal loci, at least 17 Minute loci, and at least four other haplo-abnormal loci. 3. Chromosome 3 contains one triplo-lethal locus that is also haplo-lethal, few or no other haplo-lethal loci, at least 16 Minute loci, and at least six other haplo-abnormal loci. 4. Chromosome 4 contains no triplo-lethal loci, no haplo-lethal loci, one Minute locus, and no other haplo-abnormal loci.-Thus, the Drosophila genome contains 57 loci, aneuploidy for which leads to a recognizable effect on the organism: one of these is triplo-lethal and haplo-lethal, one is triplo-abnormal and haplo-abnormal, one is hyperploid-sensitive, ten are haplo-abnormal, 41 are Minutes, and three are either haplo-lethals or Minutes. Because of the paucity of aneuploid-lethal loci, it may be concluded that the deleterious effects of aneuploidy are mostly the consequence of the additive effects of genes that are slightly sensitive to abnormal dosage. Moreover, except for the single triplo-lethal locus, the effects of hyperploidy are much less pronounced than those of the corresponding hypoploidy.     

基于红外相机技术分析鼬獾的活动节律

鼬獾(Melogale moschata)是食肉目鼬科鼬獾属动物,在我国分布广泛、种群数量丰富,但有关鼬獾的生态研究报导比较少.为掌握鼬獾的活动节律及其影响因素,2017年2月至2019年2月,利用红外相机技术对江西省桃红岭梅花鹿国家级自然保护区、九岭山国家级自然保护区和齐云山国家级自然保护区的鼬獾进行了监测,每个保护...     

Physical and biological processes influencing zonation patterns of a subtidal population of the marine snail, Astraea (Lithopoma) undosa Wood 1828

The population density and size distribution of the marine gastropod, Astraea (Lithopoma) undosa Wood 1828, at Bird Rock, Santa Catalina Island, CA, reveal an inverse relationship between population density and mean individual size, over a depth gradient. This trend may be correlated with physical and biological differences between habitats for parameters such as water motion, competitive interactions, and predation.

The potential effect of hydrodynamic forces on the zonation patterns of Astraea undosa was tested in laboratory and field experiments. Based on theoretical predictions of the relationship between shear force and water velocity on different-sized snails, large snails are subjected to greater shear forces, as a result of water motion, than medium or small snails. Results of dislodgment experiments conducted in the laboratory indicated that for a given force per unit area, all snails dislodged at nearly the same frequency, with 50% of snails predicted to dislodge at about 4 m/s, and 100% of snails predicted to dislodge at about 8 m/s velocity. These results suggest that hydrodynamic forces may be an important factor in the shallowest subtidal zones.

A factorial-designed caging experiment was used to test the effects of snail population density on growth rates of snails of three different size classes. For small and medium size classes, results indicated an inverse relationship between population density and growth rates, which was especially pronounced for smaller snails. These data, in conjunction with long-term patterns of population density and size distribution in the field, suggest that intraspecific competition also plays a role in determining size-specific zonation patterns.

Tethering experiments, used to estimate predation rates in different algal-cover zones, suggest that there are no differences in survival rates among different snail size classes; however, survival rates differ among zones and may contribute further to the observed zonation patterns. Overall, data indicate that a combination of physical and biological processes controls the population density and size-distribution of Astraea undosa over a depth gradient at Santa Catalina Island, California.     

Identification of cadherin tyrosine residues that are phosphorylated and mediate Shc association.

Previously, we reported association of the adaptor protein Shc through its SH2 domain with the cytoplasmic domain of the adhesion molecule cadherin (Xu et al. [1997] J. Biol. Chem. 272:13463-13466). This association was dependent on tyrosine phosphorylation of cadherin and could be modulated by extracellular Ca(2+) and epidermal growth factor in intact cells. There are six tyrosine residues in the cytoplasmic domain of cadherin. To define the tyrosine residue(s) that mediate Shc recognition, site-directed mutagenesis was employed to alter Tyr851 and/or Tyr883 in cadherin, which both conform to a predicted Shc SH2 domain recognition sequence. Mutation of either Tyr851 or Tyr883, but mostly the latter, decreased Src phosphorylation of cadherin and the binding of Shc to cadherin, as determined by Sepharose bead binding and gel overlay assays. Of the two tyrosine residues, Tyr883 is the major Src phosphorylation and Shc binding site. However, the double mutant (Tyr851, 883 Phe) exhibited less Shc association than the single Tyr883 Phe mutant, suggesting a role for Tyr851 also. In addition, the binding of Shc to the cadherin cytoplasmic domain was competitively inhibited by tyrosine phosphorylated peptides containing either Tyr851 or Tyr883, but not by the corresponding non-phosphorylated peptides. Mutation of Tyr851 and/or Tyr883 did not alter the capacity of the cytoplasmic domain of cadherin to bind beta-catenin in vitro. However, Shc binding to cadherin did negatively influence beta-catenin binding to the same molecule.     

Influence of different sized Daphnia species on chlorophyll concentration and summer phytoplankton community structure in eutrophic Wisconsin lakes

We conducted field observations on 13 eutrophic Wisconsin lakesdominated by either the larger bodied Daphnia pulicaria or thesmaller bodied Daphnia galeata mendotae. While Daphnia numericaldensities were not significantly different between groups oflakes, pulicaria lakes had much higher Daphnia biomasses andfiltration potentials than galeata lakes. Although we foundsignificant differences in chlorophyll (Chl) a between bothgroups of lakes during June, on a seasonal basis populationsof different sized Daphnia were not associated with significantdifferences in Chl a. Filtration potential per se was the majordeterminant of Chl a, regardless of which Daphnia species dominated.However, in pulicaria lakes, the clear-water phase started earlier,lasted longer, and was usually characterized by greater Secchidisc readings than in galeata lakes. For large blue-green algaesuch as Aphanizomenon, D.pulicaria appeared to delay bloom conditions,but ultimately did not prevent the alga from growing. Our resultssuggest that high densities of large-sized Daphnia are a desirablegoal of biomanipulation because they can attain filtration potentialshigh enough to increase summer water clarity in eutrophic lakes.     

Physiological Requirement for Both SH2 Domains for Phospholipase C-γ1 Function and Interaction with Platelet-Derived Growth Factor Receptors

Two approaches have been utilized to investigate the role of individual SH2 domains in growth factor activation of phospholipase C-gamma1 (PLC-gamma1). Surface plasmon resonance analysis indicates that the individual N-SH2 and C-SH2 domains are able to specifically recognize a phosphotyrosine-containing peptide corresponding to Tyr 1021 of the platelet-derived growth factor (PDGF) beta receptor. To assess SH2 function in the context of the full-length PLC-gamma1 molecule as well as within the intact cell, PLC-gamma1 SH2 domain mutants, disabled by site-directed mutagenesis of the N-SH2 and/or C-SH2 domain(s), were expressed in Plcg1(-/-) fibroblasts. Under equilibrium incubation conditions (4 degrees C, 40 min), the N-SH2 domain, but not the C-SH2 domain, was sufficient to mediate significant PLC-gamma1 association with the activated PDGF receptor and PLC-gamma1 tyrosine phosphorylation. When both SH2 domains in PLC-gamma1 were disabled, the double mutant did not associate with activated PDGF receptors and was not tyrosine phosphorylated. However, no single SH2 mutant was able to mediate growth factor activation of Ca2+ mobilization or inositol 1,4,5-trisphosphate (IP3) formation. Subsequent kinetic experiments demonstrated that each single SH2 domain mutant was significantly impaired in its capacity to mediate rapid association with activated PDGF receptors and become tyrosine phosphorylated. Hence, when assayed under physiological conditions necessary to achieve a rapid biological response (Ca2+ mobilization and IP3 formation), both SH2 domains of PLC-gamma1 are essential to growth factor responsiveness.     

电子供体及受体对棕色固氮菌抗氨阻遏能力的影响

电子供体连二亚硫酸钠,甲基紫精及电子受体亚甲蓝均能强烈的抑制棕色固氮菌表达固氮活性,并引起该菌的抗氨阻遏能力减弱,适当提高氧压,能提高菌体的固氮活性近15％,但过高的氧分压反而抑制菌体的固氮活性,此外,提高氢分压能降低棕色固氮菌菌体内的还原电位,从而达到提高菌体抗氨阻遏能力的效果。