NR AMZW
AU Zhou,P.; Derkatch,I.L.; Liebman,S.W.
TI The relationship between visible intracellular aggregates that appear after overexpression of Sup35 and the yeast prion-like elements [PSI(+)] and [PIN(+)]
QU Molecular Microbiology 2001 Jan; 39(1): 37-46
PT journal article
AB 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.
MH Fungal Proteins/*biosynthesis/genetics; Guanidine/pharmacology; Heat-Shock Proteins/biosynthesis; Luminescent Proteins/biosynthesis/genetics; Prions/*biosynthesis; Protein Structure, Tertiary; Recombinant Fusion Proteins/biosynthesis; Saccharomyces cerevisiae/drug effects/*ultrastructure; Support, Non-U.S. Gov't; Support, U.S. Gov't, P.H.S.
AD Laboratory for Molecular Biology, Department of Biological Sciences, University of Illinois at Chicago, Chicago, IL 60607, USA
SP englisch
PO England