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The yeast prefoldin-like URI-orthologue Bud27 associates with the RSC nucleosome remodeler and modulates transcription.

Mirón-García MC, Garrido-Godino AI, Martínez-Fernández V, Fernández-Pevida A, Cuevas-Bermúdez A, Martín-Expósito M, Chávez S, de la Cruz J, Navarro F - Nucleic Acids Res. (2014)

Bottom Line: In this work, we present evidence of Bud27 modulating RNA pol II transcription elongation.We also reveal that Bud27 associates in vivo with the Sth1 component of the chromatin remodeling complex RSC and mediates its association with RNA pol II.Our data suggest that Bud27, in addition of contributing to Rpb5 folding within the RNA polymerases, also participates in the correct assembly of other chromatin-associated protein complexes, such as RSC, thereby modulating their activity.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología Experimental, Facultad de Ciencias Experimentales, Universidad de Jaén, Paraje de las Lagunillas, s/n, 23071, Jaén, Spain.

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Lack of Bud27 affects RNA pol II occupancy and mRNA accumulation. (A) ChIP analysis for different genes in wild-type and bud27Δ cells, performed with 8WG16 antibodies, against the CTD repeat of the Rpb1 protein. (B) ChIP analysis for the GAL1 gene under galactose-induced conditions in wild-type and bud27Δ cells. As above, precipitations were performed with 8WG16 antibodies. (C) Quantitative RT-PCR analysis of mRNA levels for different genes in wild-type and bud27Δ cells at 30ºC and after a 12-h shift to 37ºC. In A and B, the fold enrichment of the indicated gene ChIP samples relative to Whole Cell Extract (WCE) samples is plotted.
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Figure 2: Lack of Bud27 affects RNA pol II occupancy and mRNA accumulation. (A) ChIP analysis for different genes in wild-type and bud27Δ cells, performed with 8WG16 antibodies, against the CTD repeat of the Rpb1 protein. (B) ChIP analysis for the GAL1 gene under galactose-induced conditions in wild-type and bud27Δ cells. As above, precipitations were performed with 8WG16 antibodies. (C) Quantitative RT-PCR analysis of mRNA levels for different genes in wild-type and bud27Δ cells at 30ºC and after a 12-h shift to 37ºC. In A and B, the fold enrichment of the indicated gene ChIP samples relative to Whole Cell Extract (WCE) samples is plotted.

Mentions: To investigate whether Bud27 participates in transcription, we first analysed the RNA pol II occupancy of a set of constitutively expressed genes (ACT1, PMA1, PYK1 and TEF2), using ChIP experiments with the 8WG16 antibody (raised against the largest subunit of the RNA pol II, Rpb1). As shown in Figure 2A, Rpb1 occupancy decreased in bud27 -mutant cells both at promoters and Open Reading Frames (ORFs), although to a different extent, depending on the gene. We also analysed the RNA pol II occupancy of the inducible gene GAL1 under activating conditions. Again, the Rpb1 ChIP signal diminished both at the promoters and the body of the gene in bud27 -mutant versus wild-type cells (Figure 2B). This decline in RNA pol II occupancy is consistent with, and likely the consequence of, the lower amount of enzyme transported to the nucleus under Bud27 deficiency (3).


The yeast prefoldin-like URI-orthologue Bud27 associates with the RSC nucleosome remodeler and modulates transcription.

Mirón-García MC, Garrido-Godino AI, Martínez-Fernández V, Fernández-Pevida A, Cuevas-Bermúdez A, Martín-Expósito M, Chávez S, de la Cruz J, Navarro F - Nucleic Acids Res. (2014)

Lack of Bud27 affects RNA pol II occupancy and mRNA accumulation. (A) ChIP analysis for different genes in wild-type and bud27Δ cells, performed with 8WG16 antibodies, against the CTD repeat of the Rpb1 protein. (B) ChIP analysis for the GAL1 gene under galactose-induced conditions in wild-type and bud27Δ cells. As above, precipitations were performed with 8WG16 antibodies. (C) Quantitative RT-PCR analysis of mRNA levels for different genes in wild-type and bud27Δ cells at 30ºC and after a 12-h shift to 37ºC. In A and B, the fold enrichment of the indicated gene ChIP samples relative to Whole Cell Extract (WCE) samples is plotted.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150788&req=5

Figure 2: Lack of Bud27 affects RNA pol II occupancy and mRNA accumulation. (A) ChIP analysis for different genes in wild-type and bud27Δ cells, performed with 8WG16 antibodies, against the CTD repeat of the Rpb1 protein. (B) ChIP analysis for the GAL1 gene under galactose-induced conditions in wild-type and bud27Δ cells. As above, precipitations were performed with 8WG16 antibodies. (C) Quantitative RT-PCR analysis of mRNA levels for different genes in wild-type and bud27Δ cells at 30ºC and after a 12-h shift to 37ºC. In A and B, the fold enrichment of the indicated gene ChIP samples relative to Whole Cell Extract (WCE) samples is plotted.
Mentions: To investigate whether Bud27 participates in transcription, we first analysed the RNA pol II occupancy of a set of constitutively expressed genes (ACT1, PMA1, PYK1 and TEF2), using ChIP experiments with the 8WG16 antibody (raised against the largest subunit of the RNA pol II, Rpb1). As shown in Figure 2A, Rpb1 occupancy decreased in bud27 -mutant cells both at promoters and Open Reading Frames (ORFs), although to a different extent, depending on the gene. We also analysed the RNA pol II occupancy of the inducible gene GAL1 under activating conditions. Again, the Rpb1 ChIP signal diminished both at the promoters and the body of the gene in bud27 -mutant versus wild-type cells (Figure 2B). This decline in RNA pol II occupancy is consistent with, and likely the consequence of, the lower amount of enzyme transported to the nucleus under Bud27 deficiency (3).

Bottom Line: In this work, we present evidence of Bud27 modulating RNA pol II transcription elongation.We also reveal that Bud27 associates in vivo with the Sth1 component of the chromatin remodeling complex RSC and mediates its association with RNA pol II.Our data suggest that Bud27, in addition of contributing to Rpb5 folding within the RNA polymerases, also participates in the correct assembly of other chromatin-associated protein complexes, such as RSC, thereby modulating their activity.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biología Experimental, Facultad de Ciencias Experimentales, Universidad de Jaén, Paraje de las Lagunillas, s/n, 23071, Jaén, Spain.

Show MeSH
Related in: MedlinePlus