Limits...
The 19S proteasome subcomplex promotes the targeting of NuA4 HAT to the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional initiation in vivo.

Uprety B, Lahudkar S, Malik S, Bhaumik SR - Nucleic Acids Res. (2011)

Bottom Line: These observations support that the 19S proteasome subcomplex enhances the targeting of co-activator at the TFIID-dependent promoter.Such an enhanced targeting of NuA4 HAT (histone acetyltransferase) promotes the recruitment of the TFIID complex for transcriptional initiation.Collectively, our data demonstrate that the 19S proteasome subcomplex enhances the targeting of NuA4 HAT to activator Rap1p at the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional stimulation, hence providing a new role of the 19S proteasome subcomplex in establishing a specific regulatory network at the TFIID-dependent promoter for productive transcriptional initiation in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Southern Illinois University-School of Medicine, Carbondale, IL 62901, USA.

ABSTRACT
Previous studies have implicated SAGA (Spt-Ada-Gcn5-acetyltransferase) and TFIID (Transcription factor-IID)-dependent mechanisms of transcriptional activation in yeast. SAGA-dependent transcriptional activation is further regulated by the 19S proteasome subcomplex. However, the role of the 19S proteasome subcomplex in transcriptional activation of the TFIID-dependent genes has not been elucidated. Therefore, we have performed a series of chromatin immunoprecipitation, mutational and transcriptional analyses at the TFIID-dependent ribosomal protein genes such as RPS5, RPL2B and RPS11B. We find that the 19S proteasome subcomplex is recruited to the promoters of these ribosomal protein genes, and promotes the association of NuA4 (Nucleosome acetyltransferase of histone H4) co-activator, but not activator Rap1p (repressor-activator protein 1). These observations support that the 19S proteasome subcomplex enhances the targeting of co-activator at the TFIID-dependent promoter. Such an enhanced targeting of NuA4 HAT (histone acetyltransferase) promotes the recruitment of the TFIID complex for transcriptional initiation. Collectively, our data demonstrate that the 19S proteasome subcomplex enhances the targeting of NuA4 HAT to activator Rap1p at the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional stimulation, hence providing a new role of the 19S proteasome subcomplex in establishing a specific regulatory network at the TFIID-dependent promoter for productive transcriptional initiation in vivo.

Show MeSH
Both 19S base and NuA4 HAT promote the recruitment of TBP (and hence transcription) at the RPL2B and RPS11B core promoters. (A) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (B) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (C) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the esa1-ts mutant and its isogenic wild-type equivalent following ts inactivation for 1 h. (D) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the esa1-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (E) Treatment of yeast cells carrying  mutation of PDR5 with MG132 (75 µM) for 2 h does not alter recruitment of TBP to the RPL2B and RPS11B core promoters. Yeast cells were grown and cross-linked as in Figure 3F.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC3300024&req=5

gkr977-F6: Both 19S base and NuA4 HAT promote the recruitment of TBP (and hence transcription) at the RPL2B and RPS11B core promoters. (A) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (B) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (C) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the esa1-ts mutant and its isogenic wild-type equivalent following ts inactivation for 1 h. (D) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the esa1-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (E) Treatment of yeast cells carrying mutation of PDR5 with MG132 (75 µM) for 2 h does not alter recruitment of TBP to the RPL2B and RPS11B core promoters. Yeast cells were grown and cross-linked as in Figure 3F.

Mentions: Although the above results implicated an important role of the 19S base in transcriptional stimulation of RPS5, it is not known whether other ribosomal protein genes are also similarly regulated by the 19S base. In view of this, we next analyzed the role of the 19S base in recruitment of TBP to the core promoters of two other ribosomal protein genes such as RPL2B and RPS11B in the rpt4-ts mutant strain and its isogenic wild-type equivalent. Interestingly, we find that the recruitment of TBP to the core promoters of these genes was significantly impaired in the rpt4-ts mutant strain (Figure 6A). Consistently, we find that the transcription of these genes was also significantly decreased in the rpt4-ts mutant strain (Figure 6B). Thus, similar to the results obtained at RPS5, we find that the 19S base facilitates the recruitment of TBP (and hence TFIID) to the core promoters of RPL2B and RPS11B to promote transcriptional initiation.Figure 6.


The 19S proteasome subcomplex promotes the targeting of NuA4 HAT to the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional initiation in vivo.

Uprety B, Lahudkar S, Malik S, Bhaumik SR - Nucleic Acids Res. (2011)

Both 19S base and NuA4 HAT promote the recruitment of TBP (and hence transcription) at the RPL2B and RPS11B core promoters. (A) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (B) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (C) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the esa1-ts mutant and its isogenic wild-type equivalent following ts inactivation for 1 h. (D) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the esa1-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (E) Treatment of yeast cells carrying  mutation of PDR5 with MG132 (75 µM) for 2 h does not alter recruitment of TBP to the RPL2B and RPS11B core promoters. Yeast cells were grown and cross-linked as in Figure 3F.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3300024&req=5

gkr977-F6: Both 19S base and NuA4 HAT promote the recruitment of TBP (and hence transcription) at the RPL2B and RPS11B core promoters. (A) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (B) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the rpt4-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (C) Analysis of recruitment of TBP to the RPL2B and RPS11B core promoters in the esa1-ts mutant and its isogenic wild-type equivalent following ts inactivation for 1 h. (D) RT–PCR analysis of RPL2B, RPS11B and ACT1 transcripts in the esa1-ts mutant and its isogenic wild-type equivalent following 1 h ts inactivation at the non-permissive temperature. (E) Treatment of yeast cells carrying mutation of PDR5 with MG132 (75 µM) for 2 h does not alter recruitment of TBP to the RPL2B and RPS11B core promoters. Yeast cells were grown and cross-linked as in Figure 3F.
Mentions: Although the above results implicated an important role of the 19S base in transcriptional stimulation of RPS5, it is not known whether other ribosomal protein genes are also similarly regulated by the 19S base. In view of this, we next analyzed the role of the 19S base in recruitment of TBP to the core promoters of two other ribosomal protein genes such as RPL2B and RPS11B in the rpt4-ts mutant strain and its isogenic wild-type equivalent. Interestingly, we find that the recruitment of TBP to the core promoters of these genes was significantly impaired in the rpt4-ts mutant strain (Figure 6A). Consistently, we find that the transcription of these genes was also significantly decreased in the rpt4-ts mutant strain (Figure 6B). Thus, similar to the results obtained at RPS5, we find that the 19S base facilitates the recruitment of TBP (and hence TFIID) to the core promoters of RPL2B and RPS11B to promote transcriptional initiation.Figure 6.

Bottom Line: These observations support that the 19S proteasome subcomplex enhances the targeting of co-activator at the TFIID-dependent promoter.Such an enhanced targeting of NuA4 HAT (histone acetyltransferase) promotes the recruitment of the TFIID complex for transcriptional initiation.Collectively, our data demonstrate that the 19S proteasome subcomplex enhances the targeting of NuA4 HAT to activator Rap1p at the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional stimulation, hence providing a new role of the 19S proteasome subcomplex in establishing a specific regulatory network at the TFIID-dependent promoter for productive transcriptional initiation in vivo.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Southern Illinois University-School of Medicine, Carbondale, IL 62901, USA.

ABSTRACT
Previous studies have implicated SAGA (Spt-Ada-Gcn5-acetyltransferase) and TFIID (Transcription factor-IID)-dependent mechanisms of transcriptional activation in yeast. SAGA-dependent transcriptional activation is further regulated by the 19S proteasome subcomplex. However, the role of the 19S proteasome subcomplex in transcriptional activation of the TFIID-dependent genes has not been elucidated. Therefore, we have performed a series of chromatin immunoprecipitation, mutational and transcriptional analyses at the TFIID-dependent ribosomal protein genes such as RPS5, RPL2B and RPS11B. We find that the 19S proteasome subcomplex is recruited to the promoters of these ribosomal protein genes, and promotes the association of NuA4 (Nucleosome acetyltransferase of histone H4) co-activator, but not activator Rap1p (repressor-activator protein 1). These observations support that the 19S proteasome subcomplex enhances the targeting of co-activator at the TFIID-dependent promoter. Such an enhanced targeting of NuA4 HAT (histone acetyltransferase) promotes the recruitment of the TFIID complex for transcriptional initiation. Collectively, our data demonstrate that the 19S proteasome subcomplex enhances the targeting of NuA4 HAT to activator Rap1p at the promoters of ribosomal protein genes to facilitate the recruitment of TFIID for transcriptional stimulation, hence providing a new role of the 19S proteasome subcomplex in establishing a specific regulatory network at the TFIID-dependent promoter for productive transcriptional initiation in vivo.

Show MeSH