Polar expression of ErbB-2/HER2 in epithelia. Bimodal regulation by Lin-7

ErbB-2/HER2 drives epithelial malignancies by forming heterodimers with growth factor receptors. The primordial invertebrate receptor is sorted to the basolateral epithelial surface by binding of the PDZ domain of Lin-7 to the receptor's tail. We show that all four human ErbBs are basolaterally expressed, even when the tail motif is absent. Mutagenesis of hLin-7 unveiled a second domain, KID, that binds to the kinase region of ErbBs. The PDZ interaction mediates stabilization of ErbB-2 at the basolateral surface. On the other hand, binding of KID is involved in initial delivery to the basolateral surface, and in its absence, unprocessed ErbB-2 molecules are diverted to the apical surface. Hence, distinct domains of Lin-7 regulate receptor delivery to and maintenance at the basolateral surface of epithelia.     

Nuclear pore protein gp210 is essential for viability in HeLa cells and Caenorhabditis elegans

Gp210 is an evolutionarily conserved membrane protein of the nuclear pore complex (NPC). We studied the phenotypes produced by RNAi-induced downregulation of gp210 in both human (HeLa) cells and Caenorhabditis elegans embryos. HeLa cell viability requires Gp210 activity. The dying cells accumulated clustered NPCs and aberrant membrane structures at the nuclear envelope, suggesting that gp210 is required directly or indirectly for nuclear pore formation and dilation as well as the anchoring or structural integrity of mature NPCs. Essential roles for gp210 were confirmed in C. elegans, where RNAi-induced reduction of gp210 caused embryonic lethality. The nuclear envelopes of embryos with reduced gp210 also had aberrant nuclear membrane structures and clustered NPCs, confirming that gp210 plays critical roles at the nuclear membrane through mechanisms that are conserved from nematodes to humans.     

Drug-induced ubiquitylation and degradation of ErbB receptor tyrosine kinases: implications for cancer therapy

Overexpression of ErbB-2/HER2 is associated with aggressive human malignancies, and therapeutic strategies targeting the oncoprotein are currently in different stages of clinical application. Tyrosine kinase inhibitors (TKIs) that block the nucleotide-binding site of the kinase are especially effective against tumors. Here we report an unexpected activity of TKIs: along with inhibition of tyrosine phosphorylation, they enhance ubiquitylation and accelerate endocytosis and subsequent intracellular destruction of ErbB-2 molecules. Especially potent is an irreversible TKI (CI-1033) that alkylates a cysteine specific to ErbB receptors. The degradative pathway stimulated by TKIs appears to be chaperone mediated, and is common to the heat shock protein 90 (Hsp90) antagonist geldanamycin and a stress-induced mechanism. In agreement with this conclusion, CI-1033 and geldanamycin additively inhibit tumor cell growth. Based upon a model for drug-induced degradation of ErbB-2, we propose a general strategy for selective destruction of oncoproteins by targeting their interaction with molecular chaperones.     

Interactions among Drosophila Nuclear Envelope Proteins Lamin, Otefin, and YA

The nuclear envelope plays many roles, including organizing nuclear structure and regulating nuclear events. Molecular associations of nuclear envelope proteins may contribute to the implementation of these functions. Lamin, otefin, and YA are the three Drosophila nuclear envelope proteins known in early embryos. We used the yeast two-hybrid system to explore the interactions between pairs of these proteins. The ubiquitous major lamina protein, lamin Dm, interacts with both otefin, a peripheral protein of the inner nuclear membrane, and YA, an essential, developmentally regulated protein of the nuclear lamina. In agreement with this interaction, lamin and otefin can be coimmunoprecipitated from the vesicle fraction of Drosophila embryos and colocalize in nuclear envelopes of Drosophila larval salivary gland nuclei. The two-hybrid system was further used to map the domains of interaction among lamin, otefin, and YA. Lamin’s rod domain interacts with the complete otefin protein, with otefin’s hydrophilic NH₂-terminal domain, and with two different fragments derived from this domain. Analogous probing of the interaction between lamin and YA showed that the lamin rod and tail plus part of its head domain are needed for interaction with full-length YA in the two-hybrid system. YA’s COOH-terminal region is necessary and sufficient for interaction with lamin. Our results suggest that interactions with lamin might mediate or stabilize the localization of otefin and YA in the nuclear lamina. They also suggest that the need for both otefin and lamin in mediating association of vesicles with chromatin might reflect the function of a protein complex that includes these two proteins.     

ESTIMATION OF CHLOROPHYLL a FROM TIME SERIES MEASUREMENTS OF HIGH SPECTRAL RESOLUTION REFLECTANCE IN AN EUTROPHIC LAKE

We acquired high spectral resolution reflectance data in Carter Lake, a eutrophic oxbow on the Iowa–Nebraska border, from April 1995 to April 1996. Chlorophyll a, total seston, sestonic organic matter, Secchi depth, and nephelometric turbidity were determined for each respective spectral measurement. Changes in algal taxonomic structure and abundance coincided with the development and senescence of a midsummer through autumn bloom of Anabaena. Taxonomic structure was more diverse in late winter and spring when Synedra sp. (diatom) and several chlorophytes and dinoflagellates were present. Overall, chlorophyll a varied from about 20 to 280 μg·L^?1, Secchi transparency from 18 to 74 cm, and seston dry weight from 11 to 48 mg·L^?1 in February and September, respectively. Particulate matter completely dominated lake water light attenuation. Dissolved organic matter had low optical activity. The most sensitive spectral feature to variation in chlorophyll a concentration was the magnitude of the scattering peak near 700 nm. The 700-nm peak correlated to chlorophyll concentration through the relationships between algal pigment absorption near 670 nm and the cell biomass and surface-related scattering signal in the near infrared. An algorithm relating the height of the 700-nm reflectance peak above a reference baseline between 670 and 850 nm to chlorophyll a was accurate and robust despite large variations in optical constituents caused by both strong seasonality in the algal system and short-term variations in seston from wind-induced sediment resuspension. The present algorithms were successfully used in other systems with different seasonality and productivity patterns. The coefficients of the models relating chlorophyll a and spectral reflectance variables appeared to be ecosystem specific: both the intercept and slope for the models in this study were moderately lower than for several other recently published results. We validated our algorithm coefficients with a second, independent dataset. The standard error for chlorophyll a prediction was ±28 μg·L^?1.     

Effects of fertilizer-induced reduction of invertebrates on reproductive success of Loggerhead Shrikes(Lanius ludovicianus)

Summary We examined the effect of spraying the common fertilizer, sodium ammonium nitrate, on cattle pastures in central Florida. Shrikes are considered good indicators of habitat quality. Hence, eight pairs of shrikes whose territories were sprayed were considered as the treatment group, and four pairs in unsprayed territories as controls. The fertilizer was sprayed on 18 June 1992, when the resident shrikes were into their second and third nests, and within 24 h the dominant bahia grass and patches of blackberry bushes became chlorotic. The eight treatment pairs expanded their territories significantly by 138.5% on average, and the smaller their initial territory, the greater the change. The total number of insects collected in the sprayed pasture in the first three weeks was extremely low compared to the number in the unsprayed pasture. In the control pairs no loss of eggs, young, or adults owing to abiotic causes was observed. However, in the treatment pairs seven eggs, two nestlings, and eight fledglings disappeared, or died from causes attributed to the spraying. In addition, seven females and one male disappeared, and one territory was abandoned. Our observations suggest the need for additional studies to examine the effects of supposedly environmentally friendly fertilizers and show the value of monitoring breeding bird populations in agricultural systems.

---

Düngerausbringung reduziert Nahrungsangebot und beeinträchtigt den Bruterfolg des Louisianawürgers(Lanius ludovicianus)

Zusammenfassung Die Auswirkungen der Ausbringung eines in den USA weithin gebräuchlichen Kalium-Ammonium-Nitrat-Düngers auf Viehweiden wurde an Louisianawürgern im Vergleich von acht betroffenen und vier nichtbetroffenen Paaren geprüft. Würger gelten allgemein als gute Indikatoren für Habitatqualität. Der Dünger wurde am 18. Juni 1992 ausgebracht, als die Würger mitten in ihrem Brutgeschäft waren. Bereits innerhalb eines Tages war die Vegetation stark geschädigt. Die von der Sprühung betroffenen Würger vergrößerten ihre Reviere um 138,5%, wobei Vögel mit vormals kleinen Revieren ihre Territorien besonders vergrößerten. Die Anzahl während der ersten drei Wochen nach der Ausbringung gesammelter Insekten war auf den besprühten Flächen im Vergleich zu den unbehandelten Weiden sehr gering. Während in von der Sprühung nicht betroffenen Bruten keinerlei auf abiotische Umstände zurückgehende Verluste an Eiern, Jungvögeln oder Altvögeln festzustellen waren, traten bei den Bruten auf den besprühten Flächen erhebliche Verluste auf. Neben dem Verlust an sieben Eiern, zwei Nestlingen und acht gerade flüggen Vögeln, verschwanden auch sieben Weibchen und ein Männchen, und ein Revier wurde verlassen. Die Beobachtungen verdeutlichen die Notwendigkeit, auch sog. umweltverträgliche Düngemittel auf ihre Wirkungen zu testen, und sie zeigen zugleich den Wert eines Umweltmonitorings mit Vögeln in der Agrarlandschaft.

     

The genomic and bioclimatic characterization of Ethiopian barley (Hordeum vulgare L.) unveils challenges and opportunities to adapt to a changing climate

The climate crisis is impacting agroecosystems and threatening food security of millions of smallholder farmers. Understanding the potential for current and future climatic adaptation of local crop agrobiodiversity may guide breeding efforts and support resilience of agriculture. Here, we combine a genomic and climatic characterization of a large collection of traditional barley varieties from Ethiopia, a staple for local smallholder farmers cropping in challenging environments. We find that the genomic diversity of barley landraces can be partially traced back to geographic and environmental diversity of the landscape. We employ a machine learning approach to model Ethiopian barley adaptation to current climate and to identify areas where its existing diversity may not be well adapted in future climate scenarios. We use this information to identify optimal trajectories of assisted migration compensating to detrimental effects of climate change, finding that Ethiopian barley diversity bears opportunities for adaptation to the climate crisis. We then characterize phenology traits in the collection in two common garden experiments in Ethiopia, using genome-wide association approaches to identify genomic loci associated with timing of flowering and maturity of the spike. We combine this information with genotype–environment associations finding that loci involved in flowering time may also explain environmental adaptation. Our data show that integrated genomic, climatic, and phenotypic characterizations of agrobiodiversity may provide breeding with actionable information to improve local adaptation in smallholder farming systems.