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A novel mechanism for target gene-specific SWI/SNF recruitment via the Snf2p N-terminus.

Weider M, Schröder A, Klebl F, Sauer N - Nucleic Acids Res. (2011)

Bottom Line: Chromatin-remodeling complexes regulate the expression of genes in all eukaryotic genomes.Here, we show that the N-terminus of Snf2p, the chromatin remodeling core unit of the SWI/SNF complex, is essential for the expression of VHT1, the gene of the plasma membrane H(+)/biotin symporter, and of BIO5, the gene of a 7-keto-8-aminopelargonic acid transporter, biotin biosynthetic precursor. chromatin immunoprecipitation (ChIP) analyses demonstrate that Vhr1p, the transcriptional regulator of VHT1 and BIO5 expression, is responsible for the targeting of Snf2p to the VHT1 promoter at low biotin.We identified an Snf2p mutant, Snf2p-R(15)C, that specifically abolishes the induction of VHT1 and BIO5 but not of other Snf2p-regulated genes, such as GAL1, SUC2 or INO1.

View Article: PubMed Central - PubMed

Affiliation: Molekulare Pflanzenphysiologie, FAU Erlangen-Nürnberg, Staudtstraße 5, D-91058 Erlangen, Germany.

ABSTRACT
Chromatin-remodeling complexes regulate the expression of genes in all eukaryotic genomes. The SWI/SNF complex of Saccharomyces cerevisiae is recruited to its target promoters via interactions with selected transcription factors. Here, we show that the N-terminus of Snf2p, the chromatin remodeling core unit of the SWI/SNF complex, is essential for the expression of VHT1, the gene of the plasma membrane H(+)/biotin symporter, and of BIO5, the gene of a 7-keto-8-aminopelargonic acid transporter, biotin biosynthetic precursor. chromatin immunoprecipitation (ChIP) analyses demonstrate that Vhr1p, the transcriptional regulator of VHT1 and BIO5 expression, is responsible for the targeting of Snf2p to the VHT1 promoter at low biotin. We identified an Snf2p mutant, Snf2p-R(15)C, that specifically abolishes the induction of VHT1 and BIO5 but not of other Snf2p-regulated genes, such as GAL1, SUC2 or INO1. We present a novel mechanism of target gene-specific SWI/SNF recruitment via Vhr1p and a conserved N-terminal Snf2p domain.

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Comparative qRT-PCR analysis of the expression of typical ySWI/SNF target genes. RT-PCR analysis of mRNA levels of SUC2, GAL1 and INO1 under inducing conditions, and of SER3 under repressing conditions. The snf2-R15C mutant strain, the Δsnf2 deletion mutant and the WT strain were first grown in YPD medium, cultures were split and incubated in the indicated media.
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Figure 7: Comparative qRT-PCR analysis of the expression of typical ySWI/SNF target genes. RT-PCR analysis of mRNA levels of SUC2, GAL1 and INO1 under inducing conditions, and of SER3 under repressing conditions. The snf2-R15C mutant strain, the Δsnf2 deletion mutant and the WT strain were first grown in YPD medium, cultures were split and incubated in the indicated media.

Mentions: The unexpected WT-like growth of the snf2-R15C mutant on YPGal and YPRaf medium or on SD medium w/o inositol pointed toward a normal, WT-like expression of the responsible ySWI/SNF target genes, GAL1, SUC2 and INO1. We tested the expression of these genes by RT-PCR in the SNF2 WT, the Δsnf2 strain and the snf2-R15C mutant that were grown on media inducing the expression of SUC2 (YPRaf), GAL1 (YPGal) or INO1 (SD medium w/o inositol). Moreover, we analyzed the expression of SER3, a gene of the serine biosynthetic pathway known to be de-repressed in Δsnf2 mutants on rich medium (25). As expected, the mRNA levels of all genes were comparable in the SNF2 WT strain and in the snf2-R15C mutant (Figure 7), whereas the Δsnf2 deletion mutant failed to induce expression of GAL1, SUC2 and INO1, and did not repress SER3.Figure 7.


A novel mechanism for target gene-specific SWI/SNF recruitment via the Snf2p N-terminus.

Weider M, Schröder A, Klebl F, Sauer N - Nucleic Acids Res. (2011)

Comparative qRT-PCR analysis of the expression of typical ySWI/SNF target genes. RT-PCR analysis of mRNA levels of SUC2, GAL1 and INO1 under inducing conditions, and of SER3 under repressing conditions. The snf2-R15C mutant strain, the Δsnf2 deletion mutant and the WT strain were first grown in YPD medium, cultures were split and incubated in the indicated media.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3105400&req=5

Figure 7: Comparative qRT-PCR analysis of the expression of typical ySWI/SNF target genes. RT-PCR analysis of mRNA levels of SUC2, GAL1 and INO1 under inducing conditions, and of SER3 under repressing conditions. The snf2-R15C mutant strain, the Δsnf2 deletion mutant and the WT strain were first grown in YPD medium, cultures were split and incubated in the indicated media.
Mentions: The unexpected WT-like growth of the snf2-R15C mutant on YPGal and YPRaf medium or on SD medium w/o inositol pointed toward a normal, WT-like expression of the responsible ySWI/SNF target genes, GAL1, SUC2 and INO1. We tested the expression of these genes by RT-PCR in the SNF2 WT, the Δsnf2 strain and the snf2-R15C mutant that were grown on media inducing the expression of SUC2 (YPRaf), GAL1 (YPGal) or INO1 (SD medium w/o inositol). Moreover, we analyzed the expression of SER3, a gene of the serine biosynthetic pathway known to be de-repressed in Δsnf2 mutants on rich medium (25). As expected, the mRNA levels of all genes were comparable in the SNF2 WT strain and in the snf2-R15C mutant (Figure 7), whereas the Δsnf2 deletion mutant failed to induce expression of GAL1, SUC2 and INO1, and did not repress SER3.Figure 7.

Bottom Line: Chromatin-remodeling complexes regulate the expression of genes in all eukaryotic genomes.Here, we show that the N-terminus of Snf2p, the chromatin remodeling core unit of the SWI/SNF complex, is essential for the expression of VHT1, the gene of the plasma membrane H(+)/biotin symporter, and of BIO5, the gene of a 7-keto-8-aminopelargonic acid transporter, biotin biosynthetic precursor. chromatin immunoprecipitation (ChIP) analyses demonstrate that Vhr1p, the transcriptional regulator of VHT1 and BIO5 expression, is responsible for the targeting of Snf2p to the VHT1 promoter at low biotin.We identified an Snf2p mutant, Snf2p-R(15)C, that specifically abolishes the induction of VHT1 and BIO5 but not of other Snf2p-regulated genes, such as GAL1, SUC2 or INO1.

View Article: PubMed Central - PubMed

Affiliation: Molekulare Pflanzenphysiologie, FAU Erlangen-Nürnberg, Staudtstraße 5, D-91058 Erlangen, Germany.

ABSTRACT
Chromatin-remodeling complexes regulate the expression of genes in all eukaryotic genomes. The SWI/SNF complex of Saccharomyces cerevisiae is recruited to its target promoters via interactions with selected transcription factors. Here, we show that the N-terminus of Snf2p, the chromatin remodeling core unit of the SWI/SNF complex, is essential for the expression of VHT1, the gene of the plasma membrane H(+)/biotin symporter, and of BIO5, the gene of a 7-keto-8-aminopelargonic acid transporter, biotin biosynthetic precursor. chromatin immunoprecipitation (ChIP) analyses demonstrate that Vhr1p, the transcriptional regulator of VHT1 and BIO5 expression, is responsible for the targeting of Snf2p to the VHT1 promoter at low biotin. We identified an Snf2p mutant, Snf2p-R(15)C, that specifically abolishes the induction of VHT1 and BIO5 but not of other Snf2p-regulated genes, such as GAL1, SUC2 or INO1. We present a novel mechanism of target gene-specific SWI/SNF recruitment via Vhr1p and a conserved N-terminal Snf2p domain.

Show MeSH
Related in: MedlinePlus