An engineered nonsense URA3 allele provides a versatile system to detect the presence, absence and appearance of the [PSI+] prion in Saccharomyces cerevisiae

Manogaran, Anita L.; Kirkland, Kathryn T.; Liebman, Susan W.

doi:10.1002/yea.1341

Cited by 22 publications

(32 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, Manogaran et al . (2006) reported a plasmid‐expressed chimeric ura3 nonsense construct made from an in‐frame fusion of the ura3 N‐terminus to an ade1‐14 segment containing a UGA nonsense allele at codon position 244 (Nakayashiki et al ., 2001).…”

Section: Resultsmentioning

confidence: 99%

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

Kurahashi

Nakamura

2007

Molecular Microbiology

View full text Add to dashboard Cite

show abstract

Section: Resultsmentioning

confidence: 99%

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

Kurahashi

Nakamura

2007

Molecular Microbiology

View full text Add to dashboard Cite

show abstract

“…We therefore employed 2 nonsense suppression reporters to determine whether functional RAC is required for suppression of spontaneous prion formation: the ade1-14 allele and the ura3-14 allele. 40 We used Luria-Delbruck fluctuation analysis to estimate [PSI C ] formation rates with 95% confidence intervals 39,41 by measuring conversion to Ade C Ura C in single colonies grown from individual [psi ¡ ] cells (Fig. 1B).…”

Section: The Rac Antagonizes Induced and Spontaneous Formation Of Thementioning

confidence: 99%

“…40 These reporters contain a premature stop codon that can be suppressed by [PSI C ] to enable growth on medium lacking adenine or uracil. W303 [psi ¡ ] Ade ¡ Ura ¡ strains (ade1-14, ura3-1) were transformed with the [PSI C ] reporter plasmid pLEU2ura3-14.…”

Section: Spontaneous Prion Formation Assaymentioning

confidence: 99%

“…W303 [psi ¡ ] Ade ¡ Ura ¡ strains (ade1-14, ura3-1) were transformed with the [PSI C ] reporter plasmid pLEU2ura3-14. 40 Transformants were resuspended in sterile H 2 O, diluted, plated on selection medium and incubated to obtain single colonies of approximately equal diameter for all strains (the fluctuation analysis relies on the total cell count being similar in all parallel cultures; the average number of cells per colony differed by <7% for WT and Dzuo1 strains). For each strain, 10 single colonies were excised from the agar per fluctuation analysis and resuspended in 100 ml sterile H 2 O.…”

Section: Spontaneous Prion Formation Assaymentioning

confidence: 99%

See 1 more Smart Citation

The ribosome-associated complex antagonizes prion formation in yeast

Amor

Castanzo

Delany

et al. 2015

Prion

View full text Add to dashboard Cite

ABSTRACT. The number of known fungal proteins capable of switching between alternative stable conformations is steadily increasing, suggesting that a prion-like mechanism may be broadly utilized as a means to propagate altered cellular states. To gain insight into the mechanisms by which cells regulate prion formation and toxicity we examined the role of the yeast ribosome-associated complex (RAC) in modulating both the formation of the [PSI C ] prion -an alternative conformer of Sup35 protein -and the toxicity of aggregation-prone polypeptides. The Hsp40 RAC chaperone Zuo1 anchors the RAC to ribosomes and stimulates the ATPase activity of the Hsp70 chaperone Ssb. We found that cells lacking Zuo1 are sensitive to over-expression of some aggregation-prone proteins, including the Sup35 prion domain, suggesting that co-translational protein misfolding increases in Dzuo1 strains. Consistent with this finding, Dzuo1 cells exhibit higher frequencies of spontaneous and induced prion formation. Cells expressing mutant forms of Zuo1 lacking either a C-terminal charged region required for ribosome association, or the J-domain responsible for Ssb ATPase stimulation, exhibit similarly high frequencies of prion formation. Our findings are consistent with a role for the RAC in chaperoning nascent Sup35 to regulate folding of the N-terminal prion domain as it emerges from the ribosome.

show abstract

“…An alternative positive selectable marker for the loss of [PSI + ] has also been developed, based on ura3-14, a nonsense allele of the URA3 gene. 40 Using such phenotypic screens, two basic classes of nuclear genetic antagonists of [PSI + ] have been identified that have arisen either spontaneously or as a consequence of chemical or UV mutagenesis (Table 1):…”

Section: Nuclear Genetic Antagonists Of [Psi + ]mentioning

confidence: 99%

The Genetic Control of the Formation and Propagation of the [PSI⁺] Prion of Yeast

Tuite

Cox

2007

Prion

View full text Add to dashboard Cite

It is over 40 years since it was first reported that the yeast Saccahromyces cerevisiae contains two unusual cytoplasmic 'genetic' elements: [PSI(+)] and [URE3]. Remarkably the underlying determinants are protein-based rather than nucleic acid-based, i.e., that they are prions, and we have already learnt much about their inheritance and phenotypic effects from the application of 'classical' genetic studies alongside the more modern molecular, cellular and biochemical approaches. Of particular value has been the exploitation of chemical mutagens and 'antagonistic' mutants which directly affect the replication and/or transmission of yeast prions. In this Chapter we describe what has emerged from the application of classical and molecular genetic studies, to the most intensively studied of the three native yeast prions, the [PSI(+)] prion.

show abstract

An engineered nonsense URA3 allele provides a versatile system to detect the presence, absence and appearance of the [PSI⁺] prion in Saccharomyces cerevisiae

Abstract: Common methods to identify yeast cells containing the prion form of the

Cited by 22 publications

References 33 publications

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

The ribosome-associated complex antagonizes prion formation in yeast

The Genetic Control of the Formation and Propagation of the [PSI⁺] Prion of Yeast

Contact Info

Product

Resources

About

An engineered nonsense URA3 allele provides a versatile system to detect the presence, absence and appearance of the [PSI+] prion in Saccharomyces cerevisiae

Abstract: Common methods to identify yeast cells containing the prion form of the

Cited by 22 publications

References 33 publications

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

Channel mutations in Hsp104 hexamer distinctively affect thermotolerance and prion‐specific propagation

The ribosome-associated complex antagonizes prion formation in yeast

The Genetic Control of the Formation and Propagation of the [PSI+] Prion of Yeast

Contact Info

Product

Resources

About

An engineered nonsense URA3 allele provides a versatile system to detect the presence, absence and appearance of the [PSI⁺] prion in Saccharomyces cerevisiae

The Genetic Control of the Formation and Propagation of the [PSI⁺] Prion of Yeast