Single-molecule imaging reveals mechanisms of protein disruption by a DNA translocase

In physiological settings, nucleic-acid translocases must act on substrates occupied by other proteins, and an increasingly appreciated role of translocases is to catalyse protein displacement from RNA and DNA. However, little is known regarding the inevitable collisions that must occur, and the fate of protein obstacles and the mechanisms by which they are evicted from DNA remain unexplored. Here we sought to establish the mechanistic basis for protein displacement from DNA using RecBCD as a model system. Using nanofabricated curtains of DNA and multicolour single-molecule microscopy, we visualized collisions between a model translocase and different DNA-bound proteins in real time. We show that the DNA translocase RecBCD can disrupt core RNA polymerase, holoenzymes, stalled elongation complexes and transcribing RNA polymerases in either head-to-head or head-to-tail orientations, as well as EcoRI(E111Q), lac repressor and even nucleosomes. RecBCD did not pause during collisions and often pushed proteins thousands of base pairs before evicting them from DNA. We conclude that RecBCD overwhelms obstacles through direct transduction of chemomechanical force with no need for specific protein-protein interactions, and that proteins can be removed from DNA through active disruption mechanisms that act on a transition state intermediate as they are pushed from one nonspecific site to the next.     

Reducing uncertainty in short-term projections: Linkage of monthly and quarterly models

This paper shows how monthly data and forecasts can be used in a systematic way to improve the predictive accuracy of a quarterly macroeconometric model. The problem is formulated as a model pooling procedure (equivalent to non-recursive Kalman filtering) where a baseline quarterly model forecast is modified through ‘add-factors’ or ‘constant adjustments’. The procedure ‘automatically’ constructs these adjustments in a covariance-minimizing fashion to reflect the revised expectation of the quarterly model's forecast errors, conditional on the monthly information set. Results obtained using Federal Reserve Board models indicate the potential for significant reduction in forecast error variance through application of these procedures.     

Breakdown of Fermi-liquid theory in a copper-oxide superconductor.

The behaviour of electrons in solids is well described by Landau's Fermi-liquid theory, which predicts that although electrons in a metal interact, they can still be treated as well defined fermions, which are called 'quasiparticles'. At low temperatures, the ability of quasiparticles to transport heat is given strictly by their ability to transport charge, as described by a universal relation known as the Wiedemann-Franz law, which hitherto no material has been known to violate. High-temperature superconductors have long been thought to fall outside the realm of Fermi-liquid theory, as suggested by several anomalous properties, but this has yet to be shown conclusively. Here we report an experimental test of the Wiedemann-Franz law in the normal state of a copper-oxide superconductor, (Pr,Ce)2CuO4, which reveals that the elementary excitations that carry heat in this material are not fermions. This is compelling evidence for the breakdown of Fermi-liquid theory in high-temperature superconductors.     

Evidence against Ha-ras-1 involvement in sporadic and familial melanoma

It was recently reported that different rare alleles at the Ha-ras-1 locus occurred at a significantly higher combined frequency in cancer patients than in an unaffected population. In particular, melanoma patients were reported to have a significantly higher frequency of such alleles. We have examined the frequency of rare Ha-ras-1 alleles in a large number of cases of sporadic melanoma. Our results indicate that the distribution of rare alleles in this population does not differ from that found in normal populations. Also, to test the hypothesis that a hereditary predisposition to melanoma could be inherited via an allele at the Ha-ras-1 locus, we examined the transmission of the segment of the short arm of chromosome 11 (11p) carrying the Ha-ras-1 locus in a number of families previously shown to exhibit a hereditary predisposition to melanoma and its precursor lesion, the dysplastic nevus syndrome (DNS). Our genetic linkage results thus obtained strongly exclude the association of a predisposition to melanoma or the precursor lesion with the inheritance of the Ha-ras-1 locus or the segment of chromosome 11 on which it is located. These results imply that hereditary predisposition to melanoma is associated with genes other than the Ha-ras-1 locus, contradicting the original suggestion of Krontiris et al., made on the basis of either an inadequate sample size or other misleading experimental factors.     

Nitrous oxide and tissue thiopental levels in rats

Zusammenfassung Es wird gezeigt, dass sowohl Gewebemenge wie auch Pentotalverteilung im Rattengewebe von Lachgas nicht beeinflusst wird.