Role of non-polyQ regions on the aggregation process by polyQ proteins into amyloid fibrils

Nine neurodegenerative disorders, referred to as polyglutamine diseases and including Huntington’s disease, are associated with the abnormal expansion of a polyglutamine tract inside nine unrelated proteins. This polyQ expansion is thought to be the major determinant in the development of neurotoxicity, triggering protein aggregation into amyloid fibrils. A large body of evidence however indicates that non-polyQ regions modulate the aggregation process triggered by polyQ expansions. The interplay between the polyQ tract and non-polyQ regions is complex and still not fully understood. In order to better understand it, we previously designed and characterized model polyQ proteins made of the beta-lactamase BlaP and a 23, 30, 55 or 79Q tract inserted in position 197 or 216. These chimeras recapitulate the aggregation properties of polyQ disease-associated proteins: there is a Q-threshold for the formation of amyloid fibrils, and above the threshold, the longer the polyQ, the faster the aggregation. Moreover, the structure of BlaP (native or unfolded) and the position of insertion of the polyQ tract (197 versus 216) influence their aggregation properties. In this work, (i) we discuss the role of the conformation of the host protein, BlaP, and of the location of the polyQ within BlaP on the different phases of amyloid fibril formation, the nucleation and elongation steps, using mainly quartz crystal microbalance (QCM), atomic force microscopy (AFM). Our results highlight a linear dependence of the polyQ length on the elongation rate whatever the insertion site and the conformation of BlaP. These two parameters however drastically influence the ability of a polyQ tract to trigger the nucleation and the elongation steps of amyloid fibril formation. (ii) Finally, we investigate the precise aggregation threshold and the modulating role of the N- and C-terminal polyQ flanking sequences in position 197 of BlaP by creating and characterizing new chimeras containing intermediate length polyQ tracts in position 197, or polyQ tracts inserted between two cleavage sites in position 197, respectively. We observe that the propensity to trigger the full process of amyloid fibril formation and its rate seems to be largely dependent on the polyQ length and on the polyQ flanking sequences. Altogether our results contribute to identify the important species and elements (polyQ or non-polyQ regions, monomers, oligomers or fibrils) during the aggregation process into amyloid fibrils to interfere with the latter associated with neurotoxicity.