Structure and conserved RNA binding of the PAZ domain

The discovery of RNA-mediated gene-silencing pathways, including RNA interference, highlights a fundamental role of short RNAs in eukaryotic gene regulation and antiviral defence. Members of the Dicer and Argonaute protein families are essential components of these RNA-silencing pathways. Notably, these two families possess an evolutionarily conserved PAZ (Piwi/Argonaute/Zwille) domain whose biochemical function is unknown. Here we report the nuclear magnetic resonance solution structure of the PAZ domain from Drosophila melanogaster Argonaute 1 (Ago1). The structure consists of a left-handed, six-stranded beta-barrel capped at one end by two alpha-helices and wrapped on one side by a distinctive appendage, which comprises a long beta-hairpin and a short alpha-helix. Using structural and biochemical analyses, we demonstrate that the PAZ domain binds a 5-nucleotide RNA with 1:1 stoichiometry. We map the RNA-binding surface to the open face of the beta-barrel, which contains amino acids conserved within the PAZ domain family, and we define the 5'-to-3' orientation of single-stranded RNA bound within that site. Furthermore, we show that PAZ domains from different human Argonaute proteins also bind RNA, establishing a conserved function for this domain.     

Involvement of receptor-interacting protein 2 in innate and adaptive immune responses

Host defences to microorganisms rely on a coordinated interplay between the innate and adaptive responses of immunity. Infection with intracellular bacteria triggers an immediate innate response requiring macrophages, neutrophils and natural killer cells, whereas subsequent activation of an adaptive response through development of T-helper subtype 1 cells (TH1) proceeds during persistent infection. To understand the physiological role of receptor-interacting protein 2 (Rip2), also known as RICK and CARDIAK, we generated mice with a targeted disruption of the gene coding for Rip2. Here we show that Rip2-deficient mice exhibit a profoundly decreased ability to defend against infection by the intracellular pathogen Listeria monocytogenes. Rip2-deficient macrophages infected with L. monocytogenes or treated with lipopolysaccharide (LPS) have decreased activation of NF-kappaB, whereas dominant negative Rip2 inhibited NF-kappaB activation mediated by Toll-like receptor 4 and Nod1. In vivo, Rip2-deficient mice were resistant to the lethal effects of LPS-induced endotoxic shock. Furthermore, Rip2 deficiency results in impaired interferon-gamma production in both TH1 and natural killer cells, attributed in part to defective interleukin-12-induced Stat4 activation. Our data reflect requirements for Rip2 in multiple pathways regulating immune and inflammatory responses.     

Palaeolithic paintings. Evolution of prehistoric cave art

Sophisticated examples of European palaeolithic parietal art can be seen in the caves of Altamira, Lascaux and Niaux near the Pyrenees, which date to the Magdalenian period (12,000-17,000 years ago), but paintings of comparable skill and complexity were created much earlier, some possibly more than 30,000 years ago. We have derived new radiocarbon dates for the drawings that decorate the Chauvet cave in Vallon-Pont-d'Arc, Ardèche, France, which confirm that even 30,000 years ago Aurignacian artists, already known as accomplished carvers, could create masterpieces comparable to the best Magdalenian art. Prehistorians, who have traditionally interpreted the evolution of prehistoric art as a steady progression from simple to more complex representations, may have to reconsider existing theories of the origins of art.     

Censored distribution techniques in analysis of toxicological data

Zusammenfassung Es werden Methoden der Dosis-Effekt-Analyse verglichen, die es erlauben, nicht mehr gemessene Werte in höheren Dosisbereichen zu schätzen. Obwohl die Daten eng beieinander liegen, ergab die vonGupta ⁵ 1952 entwickelte Methode zur Bestimmung von Mittelwerten befriedigende Schätzungen, verifiziert durch die Übereinstimmung innerhalb der Gruppen.

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Supported by N.I.H. Grants No. FR-05526 and No. C-6516, and National Air Pollution Control Administration Contract, CPA 70-17.     

Programmed population control by cell-cell communication and regulated killing

De novo engineering of gene circuits inside cells is extremely difficult, and efforts to realize predictable and robust performance must deal with noise in gene expression and variation in phenotypes between cells. Here we demonstrate that by coupling gene expression to cell survival and death using cell-cell communication, we can programme the dynamics of a population despite variability in the behaviour of individual cells. Specifically, we have built and characterized a 'population control' circuit that autonomously regulates the density of an Escherichia coli population. The cell density is broadcasted and detected by elements from a bacterial quorum-sensing system, which in turn regulate the death rate. As predicted by a simple mathematical model, the circuit can set a stable steady state in terms of cell density and gene expression that is easily tunable by varying the stability of the cell-cell communication signal. This circuit incorporates a mechanism for programmed death in response to changes in the environment, and allows us to probe the design principles of its more complex natural counterparts.