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

Abstract Rrd1p (resistance to rapamycin deletion 1) has been previously implicated in controlling transcription of rapamycin-regulated genes in response to rapamycin treatment. Intriguingly, we show here that Rrd1p associates with the coding sequence of a galactose-inducible and rapamycin non-responsive GAL1 gene, and promotes the association of RNA polymerase II with GAL1 in the absence of rapamycin treatment following transcriptional induction. Consistently, nucleosomal disassembly at GAL1 is impaired in the absence of Rrd1p, and GAL1 transcription is reduced in the Δrrd1 strain. Likewise, Rrd1p associates with the coding sequences of other rapamycin non-responsive and inducible GAL genes to promote their transcription in the absence of rapamycin treatment. Similarly, inducible, but rapamycin-responsive, non-GAL genes such as CTT1, STL1 and CUP1 are also regulated by Rrd1p. However, transcription of these inducible GAL and non-GAL genes is not altered in the absence of Rrd1p when the steady-state is reached after long transcriptional induction. 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.

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Underproduction of the Largest Subunit of RNA Polymerase II Causes Temperature Sensitivity, Slow Growth, and Inositol Auxotrophy in Saccharomyces cerevisiae

Genetics ◽

10.1093/genetics/142.3.737 ◽

1996 ◽

Vol 142 (3) ◽

pp. 737-747 ◽

Cited By ~ 7

Author(s):

Jacques Archambault ◽

David B Jansma ◽

James D Friesen

Keyword(s):

Saccharomyces Cerevisiae ◽

Steady State ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Reporter Gene ◽

Growth Medium ◽

Temperature Sensitivity ◽

Slow Growth ◽

Yeast Saccharomyces Cerevisiae ◽

Genes Encoding

Abstract In the yeast Saccharomyces cerevisiae, mutations in genes encoding subunits of RNA polymerase II (RNAPII) often give rise to a set of pleiotropic phenotypes that includes temperature sensitivity, slow growth and inositol auxotrophy. In this study, we show that these phenotypes can be brought about by a reduction in the intracellular concentration of RNAPII. Underproduction of RNAPII was achieved by expressing the gene (RPO21), encoding the largest subunit of the enzyme, from the LEU2 promoter or a weaker derivative of it, two promoters that can be repressed by the addition of leucine to the growth medium. We found that cells that underproduced RPO21 were unable to derepress fully the expression of a reporter gene under the control of the INO1 UAS. Our results indicate that temperature sensitivity, slow growth and inositol auxotrophy is a set of phenotypes that can be caused by lowering the steady-state amount of RNAPII; these results also lead to the prediction that some of the previously identified RNAPII mutations that confer this same set of phenotypes affect the assembly/stability of the enzyme. We propose a model to explain the hypersensitivity of INO1 transcription to mutations that affect components of the RNAPII transcriptional machinery.

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Heat-shock-induced variations in phosphorylation levels of the RNA polymerase II largest subunit may regulate its interaction with the peptidyl-prolyl-isomerase Pin1

Biochemistry and Cell Biology ◽

10.1139/o99-903v ◽

1999 ◽

Vol 77 (4) ◽

pp. 401

Author(s):

S Lavoie ◽

A Albert ◽

M Vincent

Keyword(s):

Heat Shock ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Prolyl Isomerase ◽

Peptidyl Prolyl Isomerase

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A heat-labile factor promotes premature 3' end formation in exon 1 of the murine adenosine deaminase gene in a cell-free transcription system

Molecular and Cellular Biology ◽

10.1128/mcb.11.11.5398-5409.1991 ◽

1991 ◽

Vol 11 (11) ◽

pp. 5398-5409

Author(s):

J W Innis ◽

R E Kellems

Keyword(s):

Steady State ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Adenosine Deaminase ◽

Free System ◽

Cell Free System ◽

Transcription System ◽

Exon 1 ◽

A Cell ◽

Processed Products

An elongation block to RNA polymerase II transcription in exon 1 is a major regulatory step in expression of the murine adenosine deaminase (ADA) gene. Previous work in the laboratory identified abundant short transcripts with 3' termini in exon 1 in steady-state RNA from injected oocytes. Using a cell-free system to investigate the mechanism of premature 3' end formation, we found that polymerase II generates prominent ADA transcripts approximately 96 to 100 nucleotides in length which are similar to the major short transcripts found in steady-state RNA from oocytes injected with ADA templates. We have determined that these transcripts are the processed products of 108- to 112-nucleotide precursors. Precursor formation is (i) favored in reactions using circular templates, (ii) not the result of a posttranscriptional processing event, (iii) sensitive to low concentrations of Sarkosyl, and (iv) dependent on a factor(s) which is inactivated in crude extracts at 47 degrees C for 15 min. The cell-free system will allow further characterization of the template and factor requirements involved in the control of premature 3' end formation by RNA polymerase II.

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Histone H3 Lysine 9 Methylation and HP1γ Are Associated with Transcription Elongation through Mammalian Chromatin.

Blood ◽

10.1182/blood.v106.11.1734.1734 ◽

2005 ◽

Vol 106 (11) ◽

pp. 1734-1734 ◽

Cited By ~ 1

Author(s):

Christopher R. Vakoc ◽

Sean A. Mandat ◽

Ben A. Olenschock ◽

Gerd A. Blobel

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Histone Modifications ◽

Transcription Elongation ◽

Gene Activation ◽

Histone H3 ◽

H3k9 Methylation ◽

Erythroid Cells ◽

Cellular Memory ◽

Active Genes

Abstract Epigenetic regulation of gene expression plays a fundamental role during tissue specification and cellular memory. Cells that are committed to a given lineage, for example during hematopoiesis, “remember” their phenotype throughout successive rounds of cell division, reflecting alterations in chromatin structure at genes that are permanently activated or silenced. Cellular memory is anchored in specific sets of histone modifications, which together form the basis for the histone code. This is illustrated in the methylation of histone molecules: while methylation of histone H3 on lysines 4, 36, and 79 is linked with gene activation, methylation of H3 on lysines 9 and 27 and histone H4 on lysine 20 is associated with transcriptionally silent heterochromatin and repressed genes within euchromatin. Not surprisingly, dysregulation of histone methylation contributes to human diseases such as leukemias. Here we examined the methylation of histone molecules during gene activation and repression triggered by the hematopoietic transcription factor GATA-1. Surprisingly, we found that during activation by GATA-1 in erythroid cells, the levels of H3K9 di- and tri-methylation increase dramatically at all examined GATA-1-stimulated genes, including alpha- and beta-globin, AHSP, Band 3 and Glycophorin A. In contrast, at all GATA-1-repressed genes examined (GATA-2, c-kit, and c-myc) these marks are rapidly lost. Peaks of H3K9 methylation were observed in the transcribed portion of genes with lower signals at the promoter regions. Heterochromatin Protein 1γ (HP1γ), a protein containing a chromo-domain that recognizes H3K9 methylation, is also present in the transcribed region of all active genes examined. We extended these analyses to include numerous genes in diverse cell types (primary erythroid cells, primary T-lymphoid cells, epithelial cells and fibroblast) and consistently found a tight correlation between H3K9 methylation and gene activity, highlighting the general nature of our findings. Both the presence of HP1γ and H3K9 methylation at active genes are dependent upon transcription elongation by RNA polymerase II. Finally, HP1γ is in a physical complex with the elongating form of RNA polymerase II. Together, our results show that H3K9 methylation and HP1γ not only function in repressive chromatin, but play a novel and unexpected role during transcription activation. These results further elucidate new combinations of histone modifications that distinguish between repressed and actively transcribing chromatin.

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Effect of hydrostatic pressure on isolated cardiac nuclei: stimulation of RNA polymerase II activity

Cardiovascular Research ◽

10.1093/cvr/12.5.265 ◽

1978 ◽

Vol 12 (5) ◽

pp. 265-268 ◽

Cited By ~ 14

Author(s):

S. S. SCHREIBER ◽

M. ORATZ ◽

M. A. ROTHSCHILD ◽

F. REFF

Keyword(s):

Hydrostatic Pressure ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Effect Of Hydrostatic Pressure ◽

Stimulation Of

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Multiple Roles for the Ess1 Prolyl Isomerase in the RNA Polymerase II Transcription Cycle

Molecular and Cellular Biology ◽

10.1128/mcb.00672-12 ◽

2012 ◽

Vol 32 (17) ◽

pp. 3594-3607 ◽

Cited By ~ 14

Author(s):

Z. Ma ◽

D. Atencio ◽

C. Barnes ◽

H. DeFiglio ◽

S. D. Hanes

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Multiple Roles ◽

Prolyl Isomerase ◽

Rna Polymerase Ii Transcription ◽

Transcription Cycle

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Pause Sites Promote Transcriptional Termination of Mammalian RNA Polymerase II

Molecular and Cellular Biology ◽

10.1128/mcb.26.10.3986-3996.2006 ◽

2006 ◽

Vol 26 (10) ◽

pp. 3986-3996 ◽

Cited By ~ 111

Author(s):

Natalia Gromak ◽

Steven West ◽

Nick J. Proudfoot

Keyword(s):

Steady State ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Rna Cleavage ◽

Actin Gene ◽

General Role ◽

Pol Ii ◽

Transcriptional Termination ◽

A Site ◽

Pause Site

ABSTRACT Polymerase II (Pol II) transcriptional termination depends on two independent genetic elements: poly(A) signals and downstream terminator sequences. The latter may either promote cotranscriptional RNA cleavage or pause elongating Pol II. We demonstrate that the previously characterized MAZ4 pause element promotes Pol II termination downstream of a poly(A) signal, dependent on both the proximity of the pause site and poly(A) signal and the strength of the poly(A) signal. The 5′→3′ exonuclease Xrn2 facilitates this pause-dependent termination by degrading the 3′ product of poly(A) site cleavage. The human β-actin gene also possesses poly(A) site proximal pause sequences, which like MAZ4 are G rich and promote transcriptional termination. Xrn2 depletion causes an increase in both steady-state RNA and Pol II levels downstream of the β-actin poly(A) site. Taken together, we provide new insights into the mechanism of pause site-mediated termination and establish a general role for the 5′→3′ exonuclease Xrn2 in Pol II termination.

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