Oxidant-induced cell-cycle delay in Saccharomyces cerevisiae: the involvement of the SWI6 transcription factor

Inhibition of mitosis by antimitotic drugs is thought to occur by destruction of microtubules, causing cells to arrest through the action of one or more mitotic checkpoints. We have patterned experiments in the yeast Saccharomyces cerevisiae after recent studies in mammalian cells that demonstrate the effectiveness of antimitotic drugs at concentrations that maintain spindle structure. We show that low concentrations of nocodazole delay cell division under the control of the previously identified mitotic checkpoint genes BUB1, BUB3, MAD1, and MAD2 and independently of BUB2. The same genes mediate the cell cycle delay induced in ctf13 mutants, limited for an essential kinetochore component. Our data suggest that a low concentration of nocodazole induces a cell cycle delay through checkpoint control that is sensitive to impaired kinetochore function. The BUB2 gene may be part of a separate checkpoint that responds to abnormal spindle structure.

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Genes that can bypass the CLN requirement for Saccharomyces cerevisiae cell cycle START

Molecular and Cellular Biology ◽

10.1128/mcb.14.3.2041-2047.1994 ◽

1994 ◽

Vol 14 (3) ◽

pp. 2041-2047

Author(s):

C B Epstein ◽

F R Cross

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Transcription Factor ◽

Copy Number ◽

Single Gene ◽

Low Copy Number ◽

Synthetically Lethal ◽

Low Expression

Cell cycle START in Saccharomyces cerevisiae requires at least one of the three CLN genes (CLN1, CLN2, or CLN3). A total of 12 mutations bypassing this requirement were found to be dominant mutations in a single gene that we named BYC1 (for bypass of CLN requirement). We also isolated a plasmid that had cln bypass activity at a low copy number; the gene responsible was distinct from BYC1 and was identical to the recently described BCK2 gene. Strains carrying bck2::ARG4 disruption alleles were fully viable, but bck2::ARG4 completely suppressed the cln bypass activity of BYC1. swi4 and swi6 deletion alleles also efficiently suppressed BYC1 cln bypass activity; Swi4 and Swi6 are components of a transcription factor previously implicated in control of CLN1 and CLN2 expression. bck2::ARG4 was synthetically lethal with cln3 deletion, suggesting that CLN1 and CLN2 cannot function in the simultaneous absence of BCK2 and CLN3; this observation correlates with low expression of CLN1 and CLN2 in bck2 strains deprived of CLN3 function. Thus, factors implicated in CLN1 and CLN2 expression and/or function are also required for BYC1 function in the absence of all three CLN genes; this may suggest the involvement of other targets of Swi4, Swi6, and Bck2 in START.

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Role of Swi4 in cell cycle regulation of CLN2 expression

Molecular and Cellular Biology ◽

10.1128/mcb.14.7.4779-4787.1994 ◽

1994 ◽

Vol 14 (7) ◽

pp. 4779-4787

Author(s):

F R Cross ◽

M Hoek ◽

J D McKinney ◽

A H Tinkelenberg

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Transcription Factor ◽

Restriction Fragment ◽

Positive Feedback ◽

Cell Cycle Regulation ◽

Transcriptional Control ◽

Regulatory Factors ◽

Cycle Regulation

Expression of the Saccharomyces cerevisiae CLN1 and CLN2 genes is cell cycle regulated, and the genes may be controlled by positive feedback. It has been proposed that positive feedback operates via Cln/Cdc28 activation of the Swi4/Swi6 transcription factor, leading to CLN1 and CLN2 transcription due to Swi4 binding to specific sites (SCBs) in the CLN1 and CLN2 promoters. To test this proposal, we have examined the effects of deletion either of the potential SCBs in the CLN2 promoter or of the SWI4 gene on CLN2 transcriptional control. Deletion of a restriction fragment containing the identified SCBs from the promoter does not prevent cell cycle regulation of CLN2 expression, although expression is lowered at all cell cycle positions. A promoter containing a 5.5-kb plasmid insertion or an independent 2.5-kb insertion at the point of deletion of the SCB-containing restriction fragment also exhibits cell cycle regulation, so involvement of unidentified upstream SCBs is unlikely. Neither Swi4 nor the related Mbp1 transcription factor is required for cell cycle regulation of the intact CLN2 promoter. In contrast, Swi4 (but not Mbp1) is required for correct cell cycle regulation of the insertion/deletion promoter lacking SCB sites. We have extended previous genetic evidence for involvement of Swi4 in some aspect of CLN2 function: a mutant hunt for CLN2 positive regulatory factors yielded only swi4 mutations at saturation. Swi4 may bind to nonconsensus sequences in the CLN2 promoter (possibly in addition to consensus sites), or it may act indirectly to regulate CLN2 expression.

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The Anticancer Ruthenium Complex KP1019 Induces DNA Damage, Leading to Cell Cycle Delay and Cell Death in Saccharomyces cerevisiae

Molecular Pharmacology ◽

10.1124/mol.112.079657 ◽

2012 ◽

Vol 83 (1) ◽

pp. 225-234 ◽

Cited By ~ 27

Author(s):

Shannon K. Stevens ◽

Amy P. Strehle ◽

Rebecca L. Miller ◽

Sarah H. Gammons ◽

Kyle J. Hoffman ◽

...

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Dna Damage ◽

Cell Death ◽

Ruthenium Complex ◽

Cell Cycle Delay

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Charcot-Marie-Tooth-related Gene GDAP1 Complements Cell Cycle Delay at G2/M Phase in Saccharomyces cerevisiae fis1 Gene-defective Cells

Journal of Biological Chemistry ◽

10.1074/jbc.m111.260042 ◽

2011 ◽

Vol 286 (42) ◽

pp. 36777-36786 ◽

Cited By ~ 20

Author(s):

Anna Estela ◽

David Pla-Martín ◽

Maribel Sánchez-Piris ◽

Hiromi Sesaki ◽

Francesc Palau

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Related Gene ◽

Charcot Marie Tooth ◽

Cell Cycle Delay ◽

M Phase

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Anatomy of a transcription factor important for the Start of the cell cycle in Saccharomyces cerevisiae

Nature ◽

10.1038/358593a0 ◽

1992 ◽

Vol 358 (6387) ◽

pp. 593-597 ◽

Cited By ~ 61

Author(s):

Michael Primig ◽

Shanthini Sockanathan ◽

Herbert Auer ◽

Kim Nasmyth

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Transcription Factor

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Telomere Cap Components Influence the Rate of Senescence in Telomerase-Deficient Yeast Cells

Molecular and Cellular Biology ◽

10.1128/mcb.24.2.837-845.2004 ◽

2004 ◽

Vol 24 (2) ◽

pp. 837-845 ◽

Cited By ~ 25

Author(s):

Shinichiro Enomoto ◽

Lynn Glowczewski ◽

Jodi Lew-Smith ◽

Judith G. Berman

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Cell Division ◽

Yeast Cells ◽

Progressive Reduction ◽

Delayed Senescence ◽

Cell Cycle Delay ◽

Nonsense Mediated Mrna Decay ◽

A Cell ◽

Population Doublings

ABSTRACT Cells lacking telomerase undergo senescence, a progressive reduction in cell division that involves a cell cycle delay and culminates in “crisis,” a period when most cells become inviable. In telomerase-deficient Saccharomyces cerevisiae cells lacking components of the nonsense-mediated mRNA decay (NMD) pathway (Upf1,Upf2, or Upf3 proteins), senescence is delayed, with crisis occurring ∼10 to 25 population doublings later than in Upf+ cells. Delayed senescence is seen in upfΔ cells lacking the telomerase holoenzyme components Est2p and TLC1 RNA, as well as in cells lacking the telomerase regulators Est1p and Est3p. The delay of senescence in upfΔ cells is not due to an increased rate of survivor formation. Rather, it is caused by alterations in the telomere cap, composed of Cdc13p, Stn1p, and Ten1p. In upfΔ mutants, STN1 and TEN1 levels are increased. Increasing the levels of Stn1p and Ten1p in Upf+ cells is sufficient to delay senescence. In addition, cdc13-2 mutants exhibit delayed senescence rates similar to those of upfΔ cells. Thus, changes in the telomere cap structure are sufficient to affect the rate of senescence in the absence of telomerase. Furthermore, the NMD pathway affects the rate of senescence in telomerase-deficient cells by altering the stoichiometry of telomere cap components.

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Role of Swi4 in cell cycle regulation of CLN2 expression.

Molecular and Cellular Biology ◽

10.1128/mcb.14.7.4779 ◽

1994 ◽

Vol 14 (7) ◽

pp. 4779-4787 ◽

Cited By ~ 41

Author(s):

F R Cross ◽

M Hoek ◽

J D McKinney ◽

A H Tinkelenberg

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Transcription Factor ◽

Restriction Fragment ◽

Positive Feedback ◽

Cell Cycle Regulation ◽

Transcriptional Control ◽

Regulatory Factors ◽

Cycle Regulation

Expression of the Saccharomyces cerevisiae CLN1 and CLN2 genes is cell cycle regulated, and the genes may be controlled by positive feedback. It has been proposed that positive feedback operates via Cln/Cdc28 activation of the Swi4/Swi6 transcription factor, leading to CLN1 and CLN2 transcription due to Swi4 binding to specific sites (SCBs) in the CLN1 and CLN2 promoters. To test this proposal, we have examined the effects of deletion either of the potential SCBs in the CLN2 promoter or of the SWI4 gene on CLN2 transcriptional control. Deletion of a restriction fragment containing the identified SCBs from the promoter does not prevent cell cycle regulation of CLN2 expression, although expression is lowered at all cell cycle positions. A promoter containing a 5.5-kb plasmid insertion or an independent 2.5-kb insertion at the point of deletion of the SCB-containing restriction fragment also exhibits cell cycle regulation, so involvement of unidentified upstream SCBs is unlikely. Neither Swi4 nor the related Mbp1 transcription factor is required for cell cycle regulation of the intact CLN2 promoter. In contrast, Swi4 (but not Mbp1) is required for correct cell cycle regulation of the insertion/deletion promoter lacking SCB sites. We have extended previous genetic evidence for involvement of Swi4 in some aspect of CLN2 function: a mutant hunt for CLN2 positive regulatory factors yielded only swi4 mutations at saturation. Swi4 may bind to nonconsensus sequences in the CLN2 promoter (possibly in addition to consensus sites), or it may act indirectly to regulate CLN2 expression.

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Ash1 and Tup1 Dependent Repression of the Saccharomyces cerevisiae HO promoter Requires Activator-Dependent Nucleosome Eviction

10.1101/2020.09.29.318063 ◽

2020 ◽

Author(s):

Emily J. Parnell ◽

Timothy J. Parnell ◽

Chao Yan ◽

Lu Bai ◽

David J. Stillman

Keyword(s):

Saccharomyces Cerevisiae ◽

Cell Cycle ◽

Transcription Factor ◽

Transcriptional Regulation ◽

Dna Sequences ◽

Binding Sites ◽

Genome Wide ◽

Intergenic Regions ◽

Mother Cells ◽

Activation Cycle

ABSTRACTTranscriptional regulation of the Saccharomyces cerevisiae HO gene is highly complex, requiring a balance of multiple activating and repressing factors to ensure that only a few transcripts are produced in mother cells within a narrow window of the cell cycle. Here, we show that the Ash1 repressor associates with two DNA sequences that are usually concealed within nucleosomes in the HO promoter and recruits the Tup1 corepressor and the Rpd3 histone deacetylase, both of which are required for full repression in daughters. Genome-wide ChIP identified greater than 200 additional sites of co-localization of these factors, primarily within large, intergenic regions from which they could regulate adjacent genes. Most Ash1 binding sites are in nucleosome depleted regions (NDRs), while a small number overlap nucleosomes, similar to HO. We demonstrate that Ash1 binding to the HO promoter does not occur in the absence of the Swi5 transcription factor, which recruits coactivators that evict nucleosomes, including the nucleosomes obscuring the Ash1 binding sites. In the absence of Swi5, artificial nucleosome depletion allowed Ash1 to bind, demonstrating that nucleosomes are inhibitory to Ash1 binding. The location of binding sites within nucleosomes may therefore be a mechanism for limiting repressive activity to periods of nucleosome eviction that are otherwise associated with activation of the promoter. Our results illustrate that activation and repression can be intricately connected, and events set in motion by an activator may also ensure the appropriate level of repression and reset the promoter for the next activation cycle.

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