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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.

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Related in: MedlinePlus

ChIP analysis of RNA polymerase II and histone H2B at GAL7 and GAL10 following transcriptional induction. (A and B) Analysis of RNA polymerase II association with the GAL7 and GAL10 coding sequence at different time points (20, 40 and 60 min) following transcriptional induction in YPG. (C and D) Analysis of RNA polymerase II levels at the GAL7 and GAL10 coding sequence following 2, 4 and 6 h transcriptional induction in YPG. (E and F) Analysis of histone H2B levels at the GAL7 and GAL10 coding sequences at different time points (30, 60 and 90 min) following transcriptional induction in YPG.
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Figure 6: ChIP analysis of RNA polymerase II and histone H2B at GAL7 and GAL10 following transcriptional induction. (A and B) Analysis of RNA polymerase II association with the GAL7 and GAL10 coding sequence at different time points (20, 40 and 60 min) following transcriptional induction in YPG. (C and D) Analysis of RNA polymerase II levels at the GAL7 and GAL10 coding sequence following 2, 4 and 6 h transcriptional induction in YPG. (E and F) Analysis of histone H2B levels at the GAL7 and GAL10 coding sequences at different time points (30, 60 and 90 min) following transcriptional induction in YPG.

Mentions: We next asked whether the effect of Rrd1p on transcription of GAL7 and GAL10 is minimal or absent when the steady-state is reached after a long induction in galactose-containing growth medium. To address this, both the wild-type and Δrrd1 strains were continuously grown in galactose-containing growth medium up to an OD600 of 1.0 prior to harvesting for RT-PCR analysis. We found that transcription of GAL7 and GAL10 in the Δrrd1 strain reached the wild-type level when the steady-state is reached after a long transcriptional induction (Figure 5H). Thus, Rrd1p promotes the initial rounds of GAL7 and GAL10 transcription, and has no effect on transcription when the steady-state is reached. This is further corroborated by the kinetic analysis of RNA polymerase II association with GAL7 and GAL10 following short or long transcriptional induction (Figure 6A–D). Moreover, the role of Rrd1p in stimulation of RNA polymerase II association with GAL7 and GAL10 is correlated with facilitated nucleosomal disassembly as the eviction of histone H2B from GAL7 and GAL10 is impaired in the Δrrd1 strain in comparison to the wild-type equivalent (Figure 6E and F).


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)

ChIP analysis of RNA polymerase II and histone H2B at GAL7 and GAL10 following transcriptional induction. (A and B) Analysis of RNA polymerase II association with the GAL7 and GAL10 coding sequence at different time points (20, 40 and 60 min) following transcriptional induction in YPG. (C and D) Analysis of RNA polymerase II levels at the GAL7 and GAL10 coding sequence following 2, 4 and 6 h transcriptional induction in YPG. (E and F) Analysis of histone H2B levels at the GAL7 and GAL10 coding sequences at different time points (30, 60 and 90 min) following transcriptional induction in YPG.
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Figure 6: ChIP analysis of RNA polymerase II and histone H2B at GAL7 and GAL10 following transcriptional induction. (A and B) Analysis of RNA polymerase II association with the GAL7 and GAL10 coding sequence at different time points (20, 40 and 60 min) following transcriptional induction in YPG. (C and D) Analysis of RNA polymerase II levels at the GAL7 and GAL10 coding sequence following 2, 4 and 6 h transcriptional induction in YPG. (E and F) Analysis of histone H2B levels at the GAL7 and GAL10 coding sequences at different time points (30, 60 and 90 min) following transcriptional induction in YPG.
Mentions: We next asked whether the effect of Rrd1p on transcription of GAL7 and GAL10 is minimal or absent when the steady-state is reached after a long induction in galactose-containing growth medium. To address this, both the wild-type and Δrrd1 strains were continuously grown in galactose-containing growth medium up to an OD600 of 1.0 prior to harvesting for RT-PCR analysis. We found that transcription of GAL7 and GAL10 in the Δrrd1 strain reached the wild-type level when the steady-state is reached after a long transcriptional induction (Figure 5H). Thus, Rrd1p promotes the initial rounds of GAL7 and GAL10 transcription, and has no effect on transcription when the steady-state is reached. This is further corroborated by the kinetic analysis of RNA polymerase II association with GAL7 and GAL10 following short or long transcriptional induction (Figure 6A–D). Moreover, the role of Rrd1p in stimulation of RNA polymerase II association with GAL7 and GAL10 is correlated with facilitated nucleosomal disassembly as the eviction of histone H2B from GAL7 and GAL10 is impaired in the Δrrd1 strain in comparison to the wild-type equivalent (Figure 6E and F).

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