A new inducible protein expression system in fission yeast based on the glucose-repressed inv1 promoter

Iacovoni, Jason S.; Russell, Paul; Gaits, Frédérique

doi:10.1016/s0378-1119(99)00116-x

Cited by 49 publications

(39 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…S1C; Iacovoni et al 1999;Choi et al 2005). Samples for ChIP-microarray analysis were taken 0 and 2 h after induction of H3-HA expression.…”

Section: Resultsmentioning

confidence: 99%

“…Locus-specific nucleosome turnover is commonly measured by the incorporation of epitope-tagged H3 that is expressed using an inducible promoter (Iacovoni et al 1999;Choi et al 2005;Aygün et al 2013;Kraushaar et al 2013). However, histone expression at nonphysiological levels may be problematic in that histone transcription is normally highly regulated in a cell cycle-dependent manner, and histone expression is extremely low, except during S phase (Rustici et al 2004).…”

mentioning

confidence: 99%

See 1 more Smart Citation

A nucleosome turnover map reveals that the stability of histone H4 Lys20 methylation depends on histone recycling in transcribed chromatin

et al. 2015

View full text Add to dashboard Cite

Nucleosome composition actively contributes to chromatin structure and accessibility. Cells have developed mechanisms to remove or recycle histones, generating a landscape of differentially aged nucleosomes. This study aimed to create a highresolution, genome-wide map of nucleosome turnover in Schizosaccharomyces pombe. The recombination-induced tag exchange (RITE) method was used to study replication-independent nucleosome turnover through the appearance of new histone H3 and the disappearance or preservation of old histone H3. The genome-wide location of histones was determined by chromatin immunoprecipitation-exonuclease methodology (ChIP-exo). The findings were compared with diverse chromatin marks, including histone variant H2A.Z, post-translational histone modifications, and Pol II binding. Finally, genomewide mapping of the methylation states of H4K20 was performed to determine the relationship between methylation (mono, di, and tri) of this residue and nucleosome turnover. Our analysis showed that histone recycling resulted in low nucleosome turnover in the coding regions of active genes, stably expressed at intermediate levels. High levels of transcription resulted in the incorporation of new histones primarily at the end of transcribed units. H4K20 was methylated in lowturnover nucleosomes in euchromatic regions, notably in the coding regions of long genes that were expressed at low levels. This transcription-dependent accumulation of histone methylation was dependent on the histone chaperone complex FACT. Our data showed that nucleosome turnover is highly dynamic in the genome and that several mechanisms are at play to either maintain or suppress stability. In particular, we found that FACT-associated transcription conserves histones by recycling them and is required for progressive H4K20 methylation.

show abstract

“…S1C; Iacovoni et al 1999;Choi et al 2005). Samples for ChIP-microarray analysis were taken 0 and 2 h after induction of H3-HA expression.…”

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 99%

A nucleosome turnover map reveals that the stability of histone H4 Lys20 methylation depends on histone recycling in transcribed chromatin

et al. 2015

View full text Add to dashboard Cite

show abstract

“…Furthermore, temperature-dependent induction activity of the truncated derivative of the nmt1 promoter was also reported recently (Kumar and Singh, 2006). In addition to the nmt1 promoter, several promoters that are also used for induced expression of heterologous genes have been developed (Bellemare et al, 2001;Erler et al, 2006;Fujita et al, 2006;Iacovoni et al, 1999). These types of inducible promoters are very useful for the maintenance of cloned genes that have harmful effects on cell growth, and timely expression to investigate the function of the gene in a specific situation.…”

Section: Introductionmentioning

confidence: 86%

A series of promoters for constitutive expression of heterologous genes in fission yeast

Matsuyama

Shirai

Yoshida

2008

Yeast

View full text Add to dashboard Cite

Inducible/repressible promoters are useful for the maintenance of toxic genes or timely expression. For ectopic expression of cloned genes in the fission yeast Schizosaccharomyces pombe, the thiamine-regulatable nmt1 promoter has been widely used, since the transcriptional activity of this promoter can be controlled by thiamine. However, this property sometimes limits a certain type of research, since the expression inevitably requires cells to be cultivated under the conditions that induce promoter activation. To allow constitutive expression of heterologous genes, we cloned three promoters of cam1 + , tif51 + and ef1a-c + . Construction of a series of vectors comprising these promoters and their introduction into the fission yeast cells demonstrated that the activity was different among these promoters but was not affected by cultured media commonly used in fission yeast. Therefore, a promoter with appropriate strength would be selectable from these promoters, depending on the genes to be expressed.

show abstract

“…To confirm this observation, Srk1 protein stability was analyzed using a different approach. We analyzed Srk1 protein expression in a ⌬sty1 strain where sty1 gene was expressed under the control of the invertase (inv1) inducible promoter (Iacovoni et al, 1999) in a plasmid bearing the "heat-inducible degron cassette" (Sanchez-Diaz et al, 2004) fused at the N-terminal of the sty1 gene (pINV1-DEGRON:Sty1-HA). A shift from glucose-to sucrose-based culture medium leads to a very rapid induction of the inv1 promoter, and therefore of Sty1 (Iacovoni et al, 1999).…”

Section: Sty1 Protein But Not Its Activity Is Essential For Srk1 Stmentioning

confidence: 99%

“…We analyzed Srk1 protein expression in a ⌬sty1 strain where sty1 gene was expressed under the control of the invertase (inv1) inducible promoter (Iacovoni et al, 1999) in a plasmid bearing the "heat-inducible degron cassette" (Sanchez-Diaz et al, 2004) fused at the N-terminal of the sty1 gene (pINV1-DEGRON:Sty1-HA). A shift from glucose-to sucrose-based culture medium leads to a very rapid induction of the inv1 promoter, and therefore of Sty1 (Iacovoni et al, 1999). In addition, the "heat-inducible degron cassette" caused Sty1 degradation by a specific ubiquitin-mediated pathway when cells were shifted from 25 to 37°C (Sanchez-Diaz et al, 2004).…”

Section: Sty1 Protein But Not Its Activity Is Essential For Srk1 Stmentioning

confidence: 99%

Activation of Srk1 by the Mitogen-activated Protein Kinase Sty1/Spc1 Precedes Its Dissociation from the Kinase and Signals Its Degradation

López-Avilés

Lambea

Moldón

et al. 2008

MBoC

View full text Add to dashboard Cite

Control of cell cycle progression by stress-activated protein kinases (SAPKs) is essential for cell adaptation to extracellular stimuli. The Schizosaccharomyces pombe SAPK Sty1/Spc1 orchestrates general changes in gene expression in response to diverse forms of cytotoxic stress. Here we show that Sty1/Spc1 is bound to its target, the Srk1 kinase, when the signaling pathway is inactive. In response to stress, Sty1/Spc1 phosphorylates Srk1 at threonine 463 of the regulatory domain, inducing both activation of Srk1 kinase, which negatively regulates cell cycle progression by inhibiting Cdc25, and dissociation of Srk1 from the SAPK, which leads to Srk1 degradation by the proteasome.

show abstract

A new inducible protein expression system in fission yeast based on the glucose-repressed inv1 promoter

Cited by 49 publications

References 12 publications

A nucleosome turnover map reveals that the stability of histone H4 Lys20 methylation depends on histone recycling in transcribed chromatin

A nucleosome turnover map reveals that the stability of histone H4 Lys20 methylation depends on histone recycling in transcribed chromatin

A series of promoters for constitutive expression of heterologous genes in fission yeast

Activation of Srk1 by the Mitogen-activated Protein Kinase Sty1/Spc1 Precedes Its Dissociation from the Kinase and Signals Its Degradation

Contact Info

Product

Resources

About