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Scc2 regulates gene expression by recruiting cohesin to the chromosome as a transcriptional activator during yeast meiosis.

Lin W, Jin H, Liu X, Hampton K, Yu HG - Mol. Biol. Cell (2011)

Bottom Line: Rec8-associated meiotic cohesin is required for the full activation of the REC8 promoter, revealing that cohesin has a positive feedback on transcriptional regulation.Finally, we provide evidence that chromosomal binding of cohesin is sufficient for target-gene activation during meiosis.Our data support a noncanonical role for cohesin as a transcriptional activator during cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.

ABSTRACT
To tether sister chromatids, a protein-loading complex, including Scc2, recruits cohesin to the chromosome at discrete loci. Cohesin facilitates the formation of a higher-order chromosome structure that could also influence gene expression. How cohesin directly regulates transcription remains to be further elucidated. We report that in budding yeast Scc2 is required for sister-chromatid cohesion during meiosis for two reasons. First, Scc2 is required for activating the expression of REC8, which encodes a meiosis-specific cohesin subunit; second, Scc2 is necessary for recruiting meiotic cohesin to the chromosome to generate sister-chromatid cohesion. Using a heterologous reporter assay, we have found that Scc2 increases the activity of its target promoters by recruiting cohesin to establish an upstream cohesin-associated region in a position-dependent manner. Rec8-associated meiotic cohesin is required for the full activation of the REC8 promoter, revealing that cohesin has a positive feedback on transcriptional regulation. Finally, we provide evidence that chromosomal binding of cohesin is sufficient for target-gene activation during meiosis. Our data support a noncanonical role for cohesin as a transcriptional activator during cell differentiation.

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Chromosome association of Scc2 and Rec8 during yeast meiosis. (A) Immunofluorescence of Scc2-GFP and Rec8'3HA (strain HY2020). Yeast cells were induced to undergo synchronous meiosis, and nuclear surface spreads were prepared and stained with GFP and HA antibodies. Two representative stages, pachytene and anaphase I, are shown. Red, Rec8'3HA; green, Scc2-GFP; blue, DAPI. Bar, 2 μm. (B) The number of chromosome-associated regions of Scc2 and Rec8 was proportional to chromosome length. Chromatin immunoprecipitation combined with microarray was used to identify Scc2 and Rec8 chromosomal binding during meiosis (strain HY1644). Inset shows the overlap of the Scc2 and Rec8 binding sites. (C) Visual representation of chromosome association profile of Scc2 and Rec8 during yeast meiosis. The entire chromosome VI is shown as a representative. The scale of the y-axis is log2 ratio of immunoprecipitation to input. SGD coordinates of chromosome VI are shown at the bottom. Red, Rec8 ChIP; black, Scc2 ChIP.
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Figure 1: Chromosome association of Scc2 and Rec8 during yeast meiosis. (A) Immunofluorescence of Scc2-GFP and Rec8'3HA (strain HY2020). Yeast cells were induced to undergo synchronous meiosis, and nuclear surface spreads were prepared and stained with GFP and HA antibodies. Two representative stages, pachytene and anaphase I, are shown. Red, Rec8'3HA; green, Scc2-GFP; blue, DAPI. Bar, 2 μm. (B) The number of chromosome-associated regions of Scc2 and Rec8 was proportional to chromosome length. Chromatin immunoprecipitation combined with microarray was used to identify Scc2 and Rec8 chromosomal binding during meiosis (strain HY1644). Inset shows the overlap of the Scc2 and Rec8 binding sites. (C) Visual representation of chromosome association profile of Scc2 and Rec8 during yeast meiosis. The entire chromosome VI is shown as a representative. The scale of the y-axis is log2 ratio of immunoprecipitation to input. SGD coordinates of chromosome VI are shown at the bottom. Red, Rec8 ChIP; black, Scc2 ChIP.

Mentions: To investigate chromosome association of Scc2 during yeast meiosis, we localized Scc2 and Rec8 on surface-spread nuclei by indirect immunofluorescence (Figure 1A). At pachytene, where homologues are paired and synapsed, Scc2, like Rec8, was localized along the length of the chromosome (Figure 1A). The Scc2 signal appeared to be less continuous but overlapped with that of Rec8 (Figure 1A, upper right). This colocalization was also evident at anaphase I, when both Scc2 and Rec8 were enriched at the centromeres that were clustered around the poles (Figure 1A). These data suggest that Scc2 largely colocalizes with cohesin on meiotic yeast chromosomes.


Scc2 regulates gene expression by recruiting cohesin to the chromosome as a transcriptional activator during yeast meiosis.

Lin W, Jin H, Liu X, Hampton K, Yu HG - Mol. Biol. Cell (2011)

Chromosome association of Scc2 and Rec8 during yeast meiosis. (A) Immunofluorescence of Scc2-GFP and Rec8'3HA (strain HY2020). Yeast cells were induced to undergo synchronous meiosis, and nuclear surface spreads were prepared and stained with GFP and HA antibodies. Two representative stages, pachytene and anaphase I, are shown. Red, Rec8'3HA; green, Scc2-GFP; blue, DAPI. Bar, 2 μm. (B) The number of chromosome-associated regions of Scc2 and Rec8 was proportional to chromosome length. Chromatin immunoprecipitation combined with microarray was used to identify Scc2 and Rec8 chromosomal binding during meiosis (strain HY1644). Inset shows the overlap of the Scc2 and Rec8 binding sites. (C) Visual representation of chromosome association profile of Scc2 and Rec8 during yeast meiosis. The entire chromosome VI is shown as a representative. The scale of the y-axis is log2 ratio of immunoprecipitation to input. SGD coordinates of chromosome VI are shown at the bottom. Red, Rec8 ChIP; black, Scc2 ChIP.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3113765&req=5

Figure 1: Chromosome association of Scc2 and Rec8 during yeast meiosis. (A) Immunofluorescence of Scc2-GFP and Rec8'3HA (strain HY2020). Yeast cells were induced to undergo synchronous meiosis, and nuclear surface spreads were prepared and stained with GFP and HA antibodies. Two representative stages, pachytene and anaphase I, are shown. Red, Rec8'3HA; green, Scc2-GFP; blue, DAPI. Bar, 2 μm. (B) The number of chromosome-associated regions of Scc2 and Rec8 was proportional to chromosome length. Chromatin immunoprecipitation combined with microarray was used to identify Scc2 and Rec8 chromosomal binding during meiosis (strain HY1644). Inset shows the overlap of the Scc2 and Rec8 binding sites. (C) Visual representation of chromosome association profile of Scc2 and Rec8 during yeast meiosis. The entire chromosome VI is shown as a representative. The scale of the y-axis is log2 ratio of immunoprecipitation to input. SGD coordinates of chromosome VI are shown at the bottom. Red, Rec8 ChIP; black, Scc2 ChIP.
Mentions: To investigate chromosome association of Scc2 during yeast meiosis, we localized Scc2 and Rec8 on surface-spread nuclei by indirect immunofluorescence (Figure 1A). At pachytene, where homologues are paired and synapsed, Scc2, like Rec8, was localized along the length of the chromosome (Figure 1A). The Scc2 signal appeared to be less continuous but overlapped with that of Rec8 (Figure 1A, upper right). This colocalization was also evident at anaphase I, when both Scc2 and Rec8 were enriched at the centromeres that were clustered around the poles (Figure 1A). These data suggest that Scc2 largely colocalizes with cohesin on meiotic yeast chromosomes.

Bottom Line: Rec8-associated meiotic cohesin is required for the full activation of the REC8 promoter, revealing that cohesin has a positive feedback on transcriptional regulation.Finally, we provide evidence that chromosomal binding of cohesin is sufficient for target-gene activation during meiosis.Our data support a noncanonical role for cohesin as a transcriptional activator during cell differentiation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biological Science, Florida State University, Tallahassee, FL 32306-4370, USA.

ABSTRACT
To tether sister chromatids, a protein-loading complex, including Scc2, recruits cohesin to the chromosome at discrete loci. Cohesin facilitates the formation of a higher-order chromosome structure that could also influence gene expression. How cohesin directly regulates transcription remains to be further elucidated. We report that in budding yeast Scc2 is required for sister-chromatid cohesion during meiosis for two reasons. First, Scc2 is required for activating the expression of REC8, which encodes a meiosis-specific cohesin subunit; second, Scc2 is necessary for recruiting meiotic cohesin to the chromosome to generate sister-chromatid cohesion. Using a heterologous reporter assay, we have found that Scc2 increases the activity of its target promoters by recruiting cohesin to establish an upstream cohesin-associated region in a position-dependent manner. Rec8-associated meiotic cohesin is required for the full activation of the REC8 promoter, revealing that cohesin has a positive feedback on transcriptional regulation. Finally, we provide evidence that chromosomal binding of cohesin is sufficient for target-gene activation during meiosis. Our data support a noncanonical role for cohesin as a transcriptional activator during cell differentiation.

Show MeSH
Related in: MedlinePlus