The relationship between visible intracellular aggregates that appear after overexpression of Sup35 and the yeast prion-like elements [PSI(+)] and [PIN(+)

Overproduced fusions of Sup35 or its prion domain with green fluorescent protein (GFP) have previously been shown to form frequent dots in PSI(+)] cells. Rare foci seen in psi(-)] cells were hypothesized to indicate the de novo induction of PSI(+)] caused by the overproduced prion domain. Here, we describe novel ring-type aggregates that also appear in psi(-)] cultures upon Sup35 overproduction and show directly that dot and ring aggregates only appear in cells that have become PSI(+)]. The formation of either type of aggregate requires PIN(+)], an element needed for the induction of PSI(+)]. Although aggregates are visible predominantly in stationary-phase cultures, PSI(+)] induction starts in exponential phase, suggesting that much smaller aggregates can also propagate PSI(+)]. Such small aggregates are probably present in PSI(+)] cells and, upon Sup35-GFP overproduction, facilitate the frequent formation of dot aggregates, but only the occasional appearance of ring aggregates. In contrast, rings are very frequent when PSI(+)] cultures, including those lacking PIN(+)], are grown in the presence of GuHCl or excess Hsp104 while overexpressing Sup35-GFP. Thus, intermediates formed during PSI(+)] curing seem to facilitate ring formation. Surprisingly, GuHCl and excess Hsp104, which are known to promote loss of PSI(+)], did not prevent the de novo induction of PSI(+)] by excess Sup35 in psi(-)]PIN(+)] strains.