Identification of genes that affect sensitivity to 5-bromodeoxyuridine in the yeast Saccharomyces cerevisiae

Fujii, Michihiko; Miki, Kensuke; Takayama, Shinichi; Ayusawa, Dai

doi:10.1007/s00438-010-0535-6

Cited by 6 publications

(3 citation statements)

References 51 publications

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“…This suggests that the J-insertion machinery may require more than one unmodified T to act, even if J is already present in DNA. These results make it unlikely that the BrdU effect on J is due to the type of chromatin alterations found by BrdU incorporation in yeast, which is highly dependent on DNA sequence (Miki et al, 2010; Fujii et al, 2010). …”

Section: Discussionmentioning

confidence: 95%

Glucosylated Hydroxymethyluracil, DNA Base J, Prevents Transcriptional Readthrough in Leishmania

Luenen

Farris

Jan

et al. 2012

Cell

150

216

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SUMMARY Some Ts in nuclear DNA of trypanosomes and Leishmania are hydroxylated and glucosylated to yield base J (β-D-glucosyl-hydroxymethyluracil). In Leishmania, about 99% of J is located in telomeric repeats. We show here that most of the remaining J is located at chromosome-internal RNA polymerase II termination sites. This internal J and telomeric J can be reduced by a knockout of J-binding protein 2 (JBP2), an enzyme involved in the first step of J biosynthesis. J levels are further reduced by growing Leishmania JBP2 knockout cells in BrdU-containing medium, resulting in cell death. The loss of internal J in JBP2 knockout cells is accompanied by massive readthrough at RNA polymerase II termination sites. The readthrough varies between transcription units but may extend over 100 kb. We conclude that J is required for proper transcription termination and infer that the absence of internal J kills Leishmania by massive readthrough of transcriptional stops.

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Section: Discussionmentioning

confidence: 95%

Glucosylated Hydroxymethyluracil, DNA Base J, Prevents Transcriptional Readthrough in Leishmania

Luenen

Farris

Jan

et al. 2012

Cell

150

216

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“…In addition, the S. cerevisiae deletion libraries are useful and widely used tools for chemical screenings to perform drug sensitivityrelated genome studies. Commonly, toxicogenomic studies employ either the haploid gene deletion collections with no gene expression (Parsons et al, 2004;Arita et al, 2009;Zhou et al, 2009;Costanzo et al, 2010;Emadi et al, 2010;Fujii et al, 2010;Stefanini et al, 2010;Kwak et al, 2011) or the heterozygous diploid collection in which gene expression is reduced (also referred to as haploinsufficiency profiling; Giaever et al, 1999;Baetz et al, 2004;Hillenmeyer et al, 2008;Bendaha et al, 2011;Hoepfner et al, 2012). The aim of these approaches is to establish chemical-genetic profiles of specific compounds such as arsenic or naphthoquinones Emadi et al, 2010) or on a larger scale to develop genetic interaction maps (Parsons et al, 2004;Costanzo et al, 2010), both of which can lead to the identification of the compound's target pathways or proteins.…”

Section: Yeast Researchmentioning

confidence: 99%

Genome-wide investigation of cellular targets and mode of action of the antifungal bacterial metabolite 2,4-diacetylphloroglucinol in Saccharomyces cerevisiae

Troppens

Dmitriev

Papkovsky

et al. 2013

FEMS Yeast Research

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Saccharomyces cerevisiae is a proven model to investigate the effects of small molecules and drugs on fungal and eukaryotic cells. In this study, the mode of action of an antifungal metabolite, 2,4-diacetylphloroglucinol (DAPG), was determined. Applying a combination of genetic and physiological approaches, it was established that this bacterial metabolite acts as a proton ionophore and dissipates the proton gradient across the mitochondrial membrane. The uncoupling of respiration and ATP synthesis ultimately leads to growth inhibition and is the primary toxic effect of DAPG. A genome-wide screen identified 154 DAPG-tolerant mutants and showed that there are many alterations in cellular metabolism that can confer at least some degree of tolerance to this uncoupler. One mutant, ydc1, was studied in some more detail as it displayed increased tolerance to both DAPG and the uncoupler carbonylcyanide m-chlorophenylhydrazone (CCCP) and appears to be unconnected to other tolerant mutant strains. Deleting YDC1 alters sphingolipid homoeostasis in the cell, and we suggest here that this may be linked to reduced drug sensitivity. Sphingolipids and their derivatives are important eukaryotic signal molecules, and the observation that altering homoeostasis may affect yeast response to metabolic uncoupling agents raises some intriguing questions for future studies.

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“…We have previously isolated the strain of the yeast Saccharomyces cerevisiae that incorporates BrdU, and shown that BrdU suppresses the growth of yeast [24]. Furthermore, we showed a possible implication of histone modifications in the regulation of the sensitivity to BrdU in yeast [25,26]. Here, we examined the roles of the histone tails in the action of BrdU, and found that a short basic domain in the N‐terminal tail of histone H2B, which is termed the HBR (histone H2B repression) domain, is involved in the action of BrdU [27].…”

Section: Introductionmentioning

confidence: 99%

The key role of a basic domain of histone H2B N‐terminal tail in the action of 5‐bromodeoxyuridine to induce cellular senescence

Watanabe

Ayusawa

et al. 2022

The FEBS Journal

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5‐Bromodeoxyuridine (BrdU), a thymidine analogue, is an interesting reagent that modulates various biological phenomena. BrdU, upon incorporation into DNA, causes destabilized nucleosome positioning which leads to changes in heterochromatin organization and gene expression in cells. We have previously shown that BrdU effectively induces cellular senescence, a phenomenon of irreversible growth arrest in mammalian cells. Identification of the mechanism of action of BrdU would provide a novel insight into the molecular mechanisms of cellular senescence. Here, we showed that a basic domain in the histone H2B N‐terminal tail, termed the HBR (histone H2B repression) domain, is involved in the action of BrdU. Notably, deletion of the HBR domain causes destabilized nucleosome positioning and derepression of gene expression, as does BrdU. We also showed that the genes up‐regulated by BrdU significantly overlapped with those by deletion of the HBR domain, the result of which suggested that BrdU and deletion of the HBR domain act in a similar way. Furthermore, we showed that decreased HBR domain function induced cellular senescence or facilitated the induction of cellular senescence. These findings indicated that the HBR domain is crucially involved in the action of BrdU, and also suggested that disordered nucleosome organization may be involved in the induction of cellular senescence.

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Identification of genes that affect sensitivity to 5-bromodeoxyuridine in the yeast Saccharomyces cerevisiae

Cited by 6 publications

References 51 publications

Glucosylated Hydroxymethyluracil, DNA Base J, Prevents Transcriptional Readthrough in Leishmania

Glucosylated Hydroxymethyluracil, DNA Base J, Prevents Transcriptional Readthrough in Leishmania

Genome-wide investigation of cellular targets and mode of action of the antifungal bacterial metabolite 2,4-diacetylphloroglucinol in Saccharomyces cerevisiae

The key role of a basic domain of histone H2B N‐terminal tail in the action of 5‐bromodeoxyuridine to induce cellular senescence

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