scholarly journals Genetic Screens to Study GAA/TTC and Inverted Repeat Instability in Saccharomyces cerevisiae

2019 ◽  
pp. 103-112
Author(s):  
Wenying Guo ◽  
Kirill S. Lobachev
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Inverted Repeat ◽  
Genetic Screens ◽  
Repeat Instability

Characterization of two telomeric DNA processing reactions in Saccharomyces cerevisiae

1988 ◽  
Vol 8 (11) ◽  
pp. 4642-4650
Author(s):  
A W Murray ◽  
T E Claus ◽  
J W Szostak
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Division ◽  
Dna Polymerase ◽  
Dna Sequences ◽  
Inverted Repeat ◽  
Telomeric Sequence ◽  
Telomeric Dna ◽  
Telomeric Sequences ◽  
Telomere Elongation

We have investigated two reactions that occur on telomeric sequences introduced into Saccharomyces cerevisiae cells by transformation. The elongation reaction added repeats of the yeast telomeric sequence C1-3A to telomeric sequences at the end of linear DNA molecules. The reaction worked on the Tetrahymena telomeric sequence C4A2 and also on the simple repeat CA. The reaction was orientation specific: it occurred only when the GT-rich strand ran 5' to 3' towards the end of the molecule. Telomere elongation occurred by non-template-directed DNA synthesis rather than any type of recombination with chromosomal telomeres, because C1-3A repeats could be added to unrelated DNA sequences between the CA-rich repeats and the terminus of the transforming DNA. The elongation reaction was very efficient, and we believe that it was responsible for maintaining an average telomere length despite incomplete replication by template-directed DNA polymerase. The resolution reaction processed a head-to-head inverted repeat of telomeric sequences into two new telomeres at a frequency of 10(-2) per cell division.

scholarly journals Characterization of two telomeric DNA processing reactions in Saccharomyces cerevisiae.

1988 ◽  
Vol 8 (11) ◽  
pp. 4642-4650 ◽  
Author(s):  
A W Murray ◽  
T E Claus ◽  
J W Szostak
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Division ◽  
Dna Polymerase ◽  
Dna Sequences ◽  
Inverted Repeat ◽  
Telomeric Sequence ◽  
Telomeric Dna ◽  
Telomeric Sequences ◽  
Telomere Elongation

We have investigated two reactions that occur on telomeric sequences introduced into Saccharomyces cerevisiae cells by transformation. The elongation reaction added repeats of the yeast telomeric sequence C1-3A to telomeric sequences at the end of linear DNA molecules. The reaction worked on the Tetrahymena telomeric sequence C4A2 and also on the simple repeat CA. The reaction was orientation specific: it occurred only when the GT-rich strand ran 5' to 3' towards the end of the molecule. Telomere elongation occurred by non-template-directed DNA synthesis rather than any type of recombination with chromosomal telomeres, because C1-3A repeats could be added to unrelated DNA sequences between the CA-rich repeats and the terminus of the transforming DNA. The elongation reaction was very efficient, and we believe that it was responsible for maintaining an average telomere length despite incomplete replication by template-directed DNA polymerase. The resolution reaction processed a head-to-head inverted repeat of telomeric sequences into two new telomeres at a frequency of 10(-2) per cell division.

scholarly journals Size Of Gene Specific Inverted Repeat - Dependent Gene Deletion In Saccharomyces cerevisiae

PLoS ONE ◽  
2013 ◽  
Vol 8 (8) ◽  
pp. e72137 ◽  
Author(s):  
Chanyuen Lim ◽  
Annette Lin Luhe ◽  
Crystal Tear JingYing ◽  
Balaji Balagurunathan ◽  
Jinchuan Wu ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Inverted Repeat ◽  
Gene Deletion

scholarly journals Retrieval of HDEL proteins is required for growth of yeast cells.

1994 ◽  
Vol 127 (1) ◽  
pp. 21-28 ◽  
Author(s):  
F M Townsley ◽  
G Frigerio ◽  
H R Pelham
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Golgi Apparatus ◽  
Yeast Cells ◽  
Genetic Screens ◽  
Inactive Form ◽  
Alpha Factor ◽  
Mutant Receptors

The ERD2 gene of Saccharomyces cerevisiae encodes the receptor which retrieves HDEL-containing containing ER proteins from the Golgi apparatus. Viable erd2 mutants have been isolated that show no obvious HDEL-dependent retention of the luminal ER protein BiP, suggesting that retrieval of HDEL proteins is not essential for growth. However, cells that lack Erd2p completely have a defective Golgi apparatus and cannot grow. This observation led to the suggestion that the receptor had a second function, possibly related to its ability to recycle from Golgi to ER. In this paper we investigate the requirements for Erd2p to support growth. We show that mutations that block its recycling also prevent growth. In addition, we show that all mutant receptors that can support growth have a residual ability to retrieve BiP, which is detectable when they are overexpressed. Mere recycling of an inactive form of the receptor, mediated by a cytoplasmic KKXX sequence, is not sufficient for growth. Furthermore, saturation of the receptor by expression of an HDEL-tagged version of pro-alpha factor inhibits growth, even of strains that do not show obvious BiP retention. We conclude that growth requires the HDEL-dependent retrieval of one or more proteins, and that these proteins can be recognized even under conditions where BiP is secreted. Genetic screens have failed to identify any one protein whose loss could account for the Erd2p requirement. Therefore, a growth may require the retention of multiple HDEL proteins in the ER, or alternatively the removal of such proteins from the Golgi apparatus.

scholarly journals A Novel Plasmid-Based Microarray Screen Identifies Suppressors of rrp6Δ in Saccharomyces cerevisiae

2006 ◽  
Vol 27 (3) ◽  
pp. 1044-1055 ◽  
Author(s):  
Katharine Abruzzi ◽  
Sylvia Denome ◽  
Jens Raabjerg Olsen ◽  
Jannie Assenholt ◽  
Line Lindegaard Haaning ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Copy Number ◽  
High Copy Number ◽  
The Novel ◽  
Genetic Screens ◽  
Gene Encoding ◽  
Mrna Metabolism ◽  
High Copy Number Plasmid ◽  
Nuclear Exosome ◽  
Microarray Analyses

ABSTRACT Genetic screens in Saccharomyces cerevisiae provide novel information about interacting genes and pathways. We screened for high-copy-number suppressors of a strain with the gene encoding the nuclear exosome component Rrp6p deleted, with either a traditional plate screen for suppressors of rrp6Δ temperature sensitivity or a novel microarray enhancer/suppressor screening (MES) strategy. MES combines DNA microarray technology with high-copy-number plasmid expression in liquid media. The plate screen and MES identified overlapping, but also different, suppressor genes. Only MES identified the novel mRNP protein Nab6p and the tRNA transporter Los1p, which could not have been identified in a traditional plate screen; both genes are toxic when overexpressed in rrp6Δ strains at 37°C. Nab6p binds poly(A)+ RNA, and the functions of Nab6p and Los1p suggest that mRNA metabolism and/or protein synthesis are growth rate limiting in rrp6Δ strains. Microarray analyses of gene expression in rrp6Δ strains and a number of suppressor strains support this hypothesis.

scholarly journals An anomalous Ty1 structure attributed to an error in reverse transcription

1986 ◽  
Vol 6 (4) ◽  
pp. 1334-1338
Author(s):  
B Errede ◽  
M Company ◽  
R Swanstrom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nucleotide Sequence ◽  
Reverse Transcription ◽  
Inverted Repeat ◽  
Repeat Structure

We have determined the nucleotide sequence of both delta elements of a Ty1 transposon inserted near the CYC7 gene in the Saccharomyces cerevisiae CYC7-H2 mutant. The upstream delta element in this Ty1 has an unusual inverted repeat structure that may have been formed by an error during reverse transcription.

scholarly journals Cytotoxicity of 1-deoxysphingolipid unraveled by genome-wide genetic screens and lipidomics in Saccharomyces cerevisiae

2019 ◽  
Vol 30 (22) ◽  
pp. 2814-2826 ◽  
Author(s):  
A. Galih Haribowo ◽  
J. Thomas Hannich ◽  
Agnès H. Michel ◽  
Márton Megyeri ◽  
Maya Schuldiner ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Cell Death ◽  
Autonomic Neuropathy ◽  
Clinical Features ◽  
Genetic Screens ◽  
Actin Organization ◽  
Genome Wide ◽  
Rule Out ◽  
Er Membrane

Hereditary sensory and autonomic neuropathy (HSAN) types IA and IC (IA/C) are caused by elevated levels of an atypical class of lipid named 1-deoxysphingolipid (DoxSL). How elevated levels of DoxSL perturb the physiology of the cell and how the perturbations lead to HSAN IA/C are largely unknown. In this study, we show that C26-1-deoxydihydroceramide (C26-DoxDHCer) is highly toxic to the cell, while C16- and C18-DoxDHCer are less toxic. Genome-wide genetic screens and lipidomics revealed the dynamics of DoxSL accumulation and DoxSL species responsible for the toxicity over the course of DoxSL accumulation. Moreover, we show that disruption of F-actin organization, alteration of mitochondrial shape, and accumulation of hydrophobic bodies by DoxSL are not sufficient to cause complete cellular failure. We found that cell death coincides with collapsed ER membrane, although we cannot rule out other possible causes of cell death. Thus, we have unraveled key principles of DoxSL cytotoxicity that may help to explain the clinical features of HSAN IA/C.

scholarly journals An anomalous Ty1 structure attributed to an error in reverse transcription.

1986 ◽  
Vol 6 (4) ◽  
pp. 1334-1338 ◽  
Author(s):  
B Errede ◽  
M Company ◽  
R Swanstrom
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Nucleotide Sequence ◽  
Reverse Transcription ◽  
Inverted Repeat ◽  
Repeat Structure

We have determined the nucleotide sequence of both delta elements of a Ty1 transposon inserted near the CYC7 gene in the Saccharomyces cerevisiae CYC7-H2 mutant. The upstream delta element in this Ty1 has an unusual inverted repeat structure that may have been formed by an error during reverse transcription.

scholarly journals Yeast MEK-dependent signal transduction: response thresholds and parameters affecting fidelity.

1995 ◽  
Vol 15 (12) ◽  
pp. 6545-6553 ◽  
Author(s):  
B Yashar ◽  
K Irie ◽  
J A Printen ◽  
B J Stevenson ◽  
G F Sprague ◽  
...  
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Signal Transduction ◽  
Signal Transduction Pathways ◽  
Cell Integrity ◽  
Genetic Screens ◽  
Dependent Signal ◽  
Response Thresholds ◽  
Inductive Signal ◽  
Full Activation ◽  
Erk Kinase

Ste7p and Mkk1p are MEK (MAPK/ERK kinase) family members that function in the mating and cell integrity signal transduction pathways in Saccharomyces cerevisiae. We selected STE7 and MKK1 mutations that stimulated their respective pathways in the absence of an inductive signal. Strikingly, serine-to-proline substitutions at analogous positions in Ste7p (position 368) and Mkk1p (position 386) were recovered by independent genetic screens. Such an outcome suggests that this substitution in other MEKs would exhibit similar properties. The Ste7p-P368 variant has higher basal enzymatic activity than Ste7p but still requires induction to reach full activation. The higher activity associated with Ste7p-P368 allows it to compensate for defects in the cell integrity pathway, but it does so only when it is overproduced or when Ste5p is missing. This behavior suggests that Ste5p, which has been proposed to be a tether for the kinases in the mating pathway, contributes to Ste7p specificity.

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