Limits...
Rrd1p, an RNA polymerase II-specific prolyl isomerase and activator of phosphoprotein phosphatase, promotes transcription independently of rapamycin response.

Sen R, Malik S, Frankland-Searby S, Uprety B, Lahudkar S, Bhaumik SR - Nucleic Acids Res. (2014)

Bottom Line: Similarly, inducible, but rapamycin-responsive, non-GAL genes such as CTT1, STL1 and CUP1 are also regulated by Rrd1p.Consistently, transcription of the constitutively active genes is not changed in the Δrrd1 strain.Taken together, our results demonstrate a new function of Rrd1p in stimulation of initial rounds of transcription, but not steady-state/constitutive transcription, of both rapamycin-responsive and non-responsive genes independently of rapamycin treatment.

View Article: PubMed Central - PubMed

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

Show MeSH

Related in: MedlinePlus

Rrd1p facilitates the PIC formation at the GAL1 promoter, and enhances transcription following 90 min transcriptional induction. (A) RT-PCR analysis of GAL1 and ADH1 mRNA levels in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. (B and C) ChIP analysis for the recruitment of TBP, Rad3p and Rpb1p to the GAL1 core promoter in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. Immunoprecipitation was performed using anti-TBP antibody (obtained from Michael R. Green, University of Massachusetts Medical School) against TBP, and anti-Myc antibody against Myc-tagged Rad3p and Myc-tagged Rpb1p. Immunoprecipitated-DNA was analyzed using the primer pair targeted to the GAL1 core promoter. (D) Western blot analysis of TBP, Rad3p and actin in the wild-type and Δrrd1 strains. Yeast strains were grown as in Figure 1B.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4150799&req=5

Figure 2: Rrd1p facilitates the PIC formation at the GAL1 promoter, and enhances transcription following 90 min transcriptional induction. (A) RT-PCR analysis of GAL1 and ADH1 mRNA levels in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. (B and C) ChIP analysis for the recruitment of TBP, Rad3p and Rpb1p to the GAL1 core promoter in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. Immunoprecipitation was performed using anti-TBP antibody (obtained from Michael R. Green, University of Massachusetts Medical School) against TBP, and anti-Myc antibody against Myc-tagged Rad3p and Myc-tagged Rpb1p. Immunoprecipitated-DNA was analyzed using the primer pair targeted to the GAL1 core promoter. (D) Western blot analysis of TBP, Rad3p and actin in the wild-type and Δrrd1 strains. Yeast strains were grown as in Figure 1B.

Mentions: Since Rrd1p associates with the coding sequence of GAL1, and promotes the association of RNA polymerase II, transcription of GAL1 would be impaired in the Δrrd1 strain. To test this, we analyzed GAL1 mRNA levels in the wild-type and Δrrd1 strains following transcriptional induction in galactose-containing growth medium. We isolated total RNAs from the wild-type and Δrrd1 strains, and then performed the RT-PCR analysis which revealed significant reduction of GAL1 mRNAs in the Δrrd1 strain (Figure 2A). As a loading control, we show that the mRNA level of a constitutively active ADH1 gene is not altered in the Δrrd1 strain in comparison to the wild-type equivalent (Figure 2A), consistent with previous studies (10) that demonstrated the dispensability of Rrd1p in regulation of transcription under vegetative growth conditions. Thus, our results demonstrate a new role of Rrd1p in promoting transcription of a rapamycin non-responsive GAL1 gene following 90 min transcriptional induction independently of rapamycin treatment (or TOR pathway).


Rrd1p, an RNA polymerase II-specific prolyl isomerase and activator of phosphoprotein phosphatase, promotes transcription independently of rapamycin response.

Sen R, Malik S, Frankland-Searby S, Uprety B, Lahudkar S, Bhaumik SR - Nucleic Acids Res. (2014)

Rrd1p facilitates the PIC formation at the GAL1 promoter, and enhances transcription following 90 min transcriptional induction. (A) RT-PCR analysis of GAL1 and ADH1 mRNA levels in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. (B and C) ChIP analysis for the recruitment of TBP, Rad3p and Rpb1p to the GAL1 core promoter in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. Immunoprecipitation was performed using anti-TBP antibody (obtained from Michael R. Green, University of Massachusetts Medical School) against TBP, and anti-Myc antibody against Myc-tagged Rad3p and Myc-tagged Rpb1p. Immunoprecipitated-DNA was analyzed using the primer pair targeted to the GAL1 core promoter. (D) Western blot analysis of TBP, Rad3p and actin in the wild-type and Δrrd1 strains. Yeast strains were grown as in Figure 1B.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150799&req=5

Figure 2: Rrd1p facilitates the PIC formation at the GAL1 promoter, and enhances transcription following 90 min transcriptional induction. (A) RT-PCR analysis of GAL1 and ADH1 mRNA levels in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. (B and C) ChIP analysis for the recruitment of TBP, Rad3p and Rpb1p to the GAL1 core promoter in the wild-type and Δrrd1 strains following 90 min transcriptional induction in YPG. Immunoprecipitation was performed using anti-TBP antibody (obtained from Michael R. Green, University of Massachusetts Medical School) against TBP, and anti-Myc antibody against Myc-tagged Rad3p and Myc-tagged Rpb1p. Immunoprecipitated-DNA was analyzed using the primer pair targeted to the GAL1 core promoter. (D) Western blot analysis of TBP, Rad3p and actin in the wild-type and Δrrd1 strains. Yeast strains were grown as in Figure 1B.
Mentions: Since Rrd1p associates with the coding sequence of GAL1, and promotes the association of RNA polymerase II, transcription of GAL1 would be impaired in the Δrrd1 strain. To test this, we analyzed GAL1 mRNA levels in the wild-type and Δrrd1 strains following transcriptional induction in galactose-containing growth medium. We isolated total RNAs from the wild-type and Δrrd1 strains, and then performed the RT-PCR analysis which revealed significant reduction of GAL1 mRNAs in the Δrrd1 strain (Figure 2A). As a loading control, we show that the mRNA level of a constitutively active ADH1 gene is not altered in the Δrrd1 strain in comparison to the wild-type equivalent (Figure 2A), consistent with previous studies (10) that demonstrated the dispensability of Rrd1p in regulation of transcription under vegetative growth conditions. Thus, our results demonstrate a new role of Rrd1p in promoting transcription of a rapamycin non-responsive GAL1 gene following 90 min transcriptional induction independently of rapamycin treatment (or TOR pathway).

Bottom Line: Similarly, inducible, but rapamycin-responsive, non-GAL genes such as CTT1, STL1 and CUP1 are also regulated by Rrd1p.Consistently, transcription of the constitutively active genes is not changed in the Δrrd1 strain.Taken together, our results demonstrate a new function of Rrd1p in stimulation of initial rounds of transcription, but not steady-state/constitutive transcription, of both rapamycin-responsive and non-responsive genes independently of rapamycin treatment.

View Article: PubMed Central - PubMed

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

Show MeSH
Related in: MedlinePlus