Mechanisms of prion protein assembly into amyloid.

The conversion of the alpha-helical, cellular isoform of the prion protein (PrP(C)) to the insoluble, beta-sheet-rich, infectious, disease-causing isoform (PrP(Sc)) is the key event in prion diseases. In an earlier study, several forms of PrP were converted into a fibrillar state by using an in vitro conversion system consisting of low concentrations of SDS and 250 mM NaCl. Here, we characterize the structure of the fibril precursor state, that is, the soluble state under fibrillization conditions. CD spectroscopy, analytical ultracentrifugation, and chemical cross-linking indicate that the precursor state exists in a monomer-dimer equilibrium of partially denatured, alpha-helical PrP, with a well defined contact site of the subunits in the dimer. Using fluorescence with thioflavin T, we monitored and quantitatively described the kinetics of seeded fibril formation, including dependence of the reaction on substrate and seed concentrations. Exponential, seed-enhanced growth can be achieved in homogeneous solution, which can be enhanced by sonication. From these data, we propose a mechanistic model of fibrillization, including the presence of several intermediate structures. These studies also provide a simplified amplification system for prions.

The error threshold.

Basic principles underlying the population dynamics of bacteria and viruses are presented, with emphasis on RNA viruses. Concepts reviewed here include fitness, mutant generation, competition, selection, sequence space and the theoretical origins of quasispecies. A "wild-type" virus is no longer viewed as the fittest type, but as the center of gravity of a population landscape. A consequence of quasispecies is the existence of an error threshold for selective competence. The error threshold has a justification in quasispecies theory and lends itself to exploration of strategies to drive viral populations over the error threshold, the central theme of this volume.

Transferability of HIV by arthropods supports the hypothesis about transmission of the virus from apes to man.

The primate Pan troglodytes troglodytes, a chimpanzee subspecies, has recently been defined as a natural animal host of the human immunodeficiency virus (HIV). Apes are traditionally hunted in Africa and are offered for sale in open-air meat markets. The bloody carcasses are regularly covered with blood-feeding flies, amongst them possibly the stable fly (Stomoxvs calcitrans L.). a cosmopolitically occurring biting fly. This fly is the effective vector for the retrovirus causing equine infectious anemia [corrected]. According to laboratory experiments, the infectivity of ingested HIV is not reduced in the regurgitates of this fly. These findings are combined to explain the mechanism for a possible primary transmission of HIV from ape to man.