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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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Growth test on SD medium with varying biotin concentrations. The Δsnf2 mutant was transformed with plasmid pVHT1oe, expressing VHT1 from the constitutive ADH1 promoter (Δsnf2 + VHT1oe). To allow growth on the same medium, the deletion mutants Δvht1 and Δsnf2 and the WT were transformed with the empty vector NEV-E. Cells were spotted on SD medium containing the indicted biotin concentrations after serial 1:8 dilutions, grown for 3 days and photographed.
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Figure 5: Growth test on SD medium with varying biotin concentrations. The Δsnf2 mutant was transformed with plasmid pVHT1oe, expressing VHT1 from the constitutive ADH1 promoter (Δsnf2 + VHT1oe). To allow growth on the same medium, the deletion mutants Δvht1 and Δsnf2 and the WT were transformed with the empty vector NEV-E. Cells were spotted on SD medium containing the indicted biotin concentrations after serial 1:8 dilutions, grown for 3 days and photographed.

Mentions: Based on the data presented so far (Figures 1B, 3 and 4), we expected that snf2 mutants should be severely affected in their capacity to grow on low biotin. We, therefore, compared the growth of the Δsnf2 mutant, of the corresponding SNF2 WT and of a Δvht1 deletion mutant [JSYΔvht1 (26)] on SD media with decreasing biotin concentrations. As expected, the Δvht1 mutant did grow only on high biotin (2 mg biotin l−1; Figure 5). Also the Δsnf2 cells failed to grow on medium with low biotin (0.02 µg biotin l−1; Figure 5). The growth difference between Δvht1 and Δsnf2 cells on medium biotin concentrations (200 µg biotin l−1) results from a weak basal activity of pVHT1 in Δsnf2 cells and the complete absence of Vht1p activity in Δvht1 cells.Figure 5.


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)

Growth test on SD medium with varying biotin concentrations. The Δsnf2 mutant was transformed with plasmid pVHT1oe, expressing VHT1 from the constitutive ADH1 promoter (Δsnf2 + VHT1oe). To allow growth on the same medium, the deletion mutants Δvht1 and Δsnf2 and the WT were transformed with the empty vector NEV-E. Cells were spotted on SD medium containing the indicted biotin concentrations after serial 1:8 dilutions, grown for 3 days and photographed.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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

Figure 5: Growth test on SD medium with varying biotin concentrations. The Δsnf2 mutant was transformed with plasmid pVHT1oe, expressing VHT1 from the constitutive ADH1 promoter (Δsnf2 + VHT1oe). To allow growth on the same medium, the deletion mutants Δvht1 and Δsnf2 and the WT were transformed with the empty vector NEV-E. Cells were spotted on SD medium containing the indicted biotin concentrations after serial 1:8 dilutions, grown for 3 days and photographed.
Mentions: Based on the data presented so far (Figures 1B, 3 and 4), we expected that snf2 mutants should be severely affected in their capacity to grow on low biotin. We, therefore, compared the growth of the Δsnf2 mutant, of the corresponding SNF2 WT and of a Δvht1 deletion mutant [JSYΔvht1 (26)] on SD media with decreasing biotin concentrations. As expected, the Δvht1 mutant did grow only on high biotin (2 mg biotin l−1; Figure 5). Also the Δsnf2 cells failed to grow on medium with low biotin (0.02 µg biotin l−1; Figure 5). The growth difference between Δvht1 and Δsnf2 cells on medium biotin concentrations (200 µg biotin l−1) results from a weak basal activity of pVHT1 in Δsnf2 cells and the complete absence of Vht1p activity in Δvht1 cells.Figure 5.

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