The Coupling of Transcription Termination by RNA Polymerase II to MRNA 3' End Processing in Saccharomyces Cerevisiae

The Coupling of Transcription Termination by RNA Polymerase II to MRNA 3' End Processing in Saccharomyces Cerevisiae PDF Author: Weifei Luo
Publisher:
ISBN: 9781109838848
Category :
Languages : en
Pages : 145

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Book Description
Transcription termination by RNA polymerase II (pol II) is a process that involves the release of polymerase and the RNA from the transcription elongation complex. In eukaryotes, transcription termination is tightly coupled to mRNA 3' end processing (cleavage/polyadenylation) largely via the C-terminal domain (CTD) of pol II. The torpedo model for pol II transcription termination proposes that a 5'-3' RNA exonuclease enters at the poly (A) cleavage site, degrades the 3' nascent RNA and eventually displaces polymerase from the DNA. We directly demonstrated that two exonucleases, Rat1 and Xrn1, both contribute to co-transcriptional degradation of nascent RNA but this degradation is not sufficient to cause polymerase release. Instead, Rat1 functions in both 3' end processing and termination by enhancing recruitment of 3' end processing factors. In addition, the cleavage factor Pcf11 reciprocally aids in recruitment of Rat1 to the elongation complex. How 3' end cleavage of pre-mRNA is involved in termination is still not well understood. We introduced a variant of hepatitis delta ribozyme either in the coding region or 3' flanking region of ADI-4 gene and investigated whether severing the nascent by the self-cleaving ribozyme could affect transcription termination. We showed that ribozyme cleavage within the coding region of ADH4 stimulates premature termination and dramatically enhances the recruitment of Rat1, Pcf11 and Rna15 near the ribozyme sequence although the ribozyme inserted downstream of the ADH4 poly (A) site did not affect normal termination and replacement of the ADH4 3' UTR by the ribozyme did not restore termination in the region immediately downstream. We also demonstrated that Ctk1, Ess1 and Spt5 are required for transcription termination; and Ess1 is essential for proper recruitment of Spt5 and Pcf11 of the CFIA complex at the 3' end of a gene. These observations imply that both the CTD phosphorylation and conformational modification are important to stimulate efficient termination. In summary, the results presented in this thesis suggest a unified model for pol II transcription termination: multiple contacts among the polymerase, Rat1, CFIA complex and the RNA function together to ensure efficient termination.

The Coupling of Transcription Termination by RNA Polymerase II to MRNA 3' End Processing in Saccharomyces Cerevisiae

The Coupling of Transcription Termination by RNA Polymerase II to MRNA 3' End Processing in Saccharomyces Cerevisiae PDF Author: Weifei Luo
Publisher:
ISBN: 9781109838848
Category :
Languages : en
Pages : 145

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Book Description
Transcription termination by RNA polymerase II (pol II) is a process that involves the release of polymerase and the RNA from the transcription elongation complex. In eukaryotes, transcription termination is tightly coupled to mRNA 3' end processing (cleavage/polyadenylation) largely via the C-terminal domain (CTD) of pol II. The torpedo model for pol II transcription termination proposes that a 5'-3' RNA exonuclease enters at the poly (A) cleavage site, degrades the 3' nascent RNA and eventually displaces polymerase from the DNA. We directly demonstrated that two exonucleases, Rat1 and Xrn1, both contribute to co-transcriptional degradation of nascent RNA but this degradation is not sufficient to cause polymerase release. Instead, Rat1 functions in both 3' end processing and termination by enhancing recruitment of 3' end processing factors. In addition, the cleavage factor Pcf11 reciprocally aids in recruitment of Rat1 to the elongation complex. How 3' end cleavage of pre-mRNA is involved in termination is still not well understood. We introduced a variant of hepatitis delta ribozyme either in the coding region or 3' flanking region of ADI-4 gene and investigated whether severing the nascent by the self-cleaving ribozyme could affect transcription termination. We showed that ribozyme cleavage within the coding region of ADH4 stimulates premature termination and dramatically enhances the recruitment of Rat1, Pcf11 and Rna15 near the ribozyme sequence although the ribozyme inserted downstream of the ADH4 poly (A) site did not affect normal termination and replacement of the ADH4 3' UTR by the ribozyme did not restore termination in the region immediately downstream. We also demonstrated that Ctk1, Ess1 and Spt5 are required for transcription termination; and Ess1 is essential for proper recruitment of Spt5 and Pcf11 of the CFIA complex at the 3' end of a gene. These observations imply that both the CTD phosphorylation and conformational modification are important to stimulate efficient termination. In summary, the results presented in this thesis suggest a unified model for pol II transcription termination: multiple contacts among the polymerase, Rat1, CFIA complex and the RNA function together to ensure efficient termination.

3' Processing of Messenger RNA in the Yeast Saccharomyces Cerevisiae

3' Processing of Messenger RNA in the Yeast Saccharomyces Cerevisiae PDF Author: Christoph Mathias Egli
Publisher:
ISBN:
Category : Genetic transcription
Languages : en
Pages : 124

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Book Description


Evidence for Coupling Transcription and Splicing in Vivo in Saccharomyces Cerevisiae

Evidence for Coupling Transcription and Splicing in Vivo in Saccharomyces Cerevisiae PDF Author: Luh Tung
Publisher:
ISBN:
Category : Adenosine triphosphatase
Languages : en
Pages : 191

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Book Description
Abstract: The genetic information transfer pathway, which includes transcription, precursor messenger RNA (pre-mRNA) processing, mRNA export, translation, and mRNA turnover, is a highly integrated process in higher eukaryotes. This integration is mainly achieved by the C-terminal domain (CTD) of the largest subunit of the RNA polymerase II (Pol II), which serves as a platform for recruiting RNA processing factors. In yeast Saccharomyces cerevisiae, however, support for the coupling of transcription and splicing is far less assuring. Here we show, via our studies of Sub2p, a DExD/H-box protein essential for both splicing and mRNA export, that transcription and splicing are functionally coupled in yeast. The DExD/H-box proteins are often referred to as RNA helicases, but are increasingly viewed as ribonucleoprotein ATPases (RNPases) that can remodel specific RNPs in vivo. We first show that specific alterations of the intron branch-site binding protein (BBP) can eliminate the requirement for Sub2p, thus illuminating a principal role for Sub2p to remove BBP and Mud2p from the branch-site region. Unexpectedly, we uncovered a panoply of genetic and chemical perturbations of the yeast transcription machinery that can also bypass Sub2p's requirement. Chromatin-immunoprecipitation (ChIP) analysis revealed that these perturbations significantly reduce the effectiveness of co-transcriptional recruitment of BBP, thereby alleviating the lethal outcome of losing Sub2p. These findings also suggest a potential selective advantage for yeast to exploit modifications in a kinetically coupled upstream pathway to offset the catastrophic loss of key components in the downstream pathways.

RNA Binding to RNA Polymerase II Purified from the Yeast Saccharomyces Cerevisiae

RNA Binding to RNA Polymerase II Purified from the Yeast Saccharomyces Cerevisiae PDF Author: Tracy Lanise Johnson
Publisher:
ISBN:
Category :
Languages : en
Pages : 400

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Book Description


RNA Processing

RNA Processing PDF Author: Paula Grabowski
Publisher: BoD – Books on Demand
ISBN: 9533075570
Category : Medical
Languages : en
Pages : 262

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Book Description
RNA functions broadly as informational molecule, genome, enzyme and machinery for RNA processing. While these functions reflect ancient activities, they also remain vital components of contemporary biochemical pathways. In eukaryotic cells RNA processing impacts the biogenesis of RNA molecules of essentially every shape and function. The collection of articles in this volume describes the current state of understanding of the broad array of RNA processing events in animal and plant cells, key unanswered questions, and cutting edge approaches available to address these questions. Some questions discussed in this volume include, how viruses subvert the RNA processing machinery of the host cell, how the coordination of co-transcriptional RNA processing is regulated at the level of chromatin, the status of RNA processing in plant organelles, and how micro RNA machinery is biosynthesized and regulated.

Gene Regulation, Epigenetics and Hormone Signaling

Gene Regulation, Epigenetics and Hormone Signaling PDF Author: Subhrangsu S. Mandal
Publisher: John Wiley & Sons
ISBN: 3527697241
Category : Science
Languages : en
Pages : 688

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Book Description
Das erste Referenzwerk dieser Art mit einer umfassenden und dennoch prägnanten Einführung in die Epigenetik beschäftigt sich mit den unzähligen Interaktionen zwischen Hormonregulation und Epigenetik. Die Inhalte sind gut strukturiert. Es gibt keine Überschneidungen zwischen den Kapiteln und jedes Kapitel beinhaltet Zusatzmaterialien für Präsentationen. Der Schwerpunkt liegt durchgängig auf Erkrankungen. Zielgruppe sind die vielen Physiologen und Entwicklungsbiologen, die zwar mit der Bedeutung und den Mechanismen der Hormonregulation vertraut sind, aber über unzureichendes Hintergrundwissen im Bereich Epigenetik verfügen.

Transcription and Splicing

Transcription and Splicing PDF Author: B. D. Hames
Publisher: Oxford University Press, USA
ISBN:
Category : Music
Languages : en
Pages : 238

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Book Description
This book gives a co-ordinated review of our present knowledge of eukaryotic RNA synthesis.

Biochemical and Genetic Characterization of the Transcription Elongation Factor TFIIS from the Yeast Saccharomyces Cerevisiae

Biochemical and Genetic Characterization of the Transcription Elongation Factor TFIIS from the Yeast Saccharomyces Cerevisiae PDF Author: Karen Renee Christie
Publisher:
ISBN:
Category :
Languages : en
Pages : 600

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Book Description
Regulation of the process of transcriptional elongation is an important control mechanism in the expression of some genes. To fully understand this form of regulation will require better understanding of the functions of transcription elongation factors. The goal of this work was to characterize the transcription elongation factor TFIIS from Saccharomyces cerevisiae, originally called P37. I demonstrated that, like the mammalian TFIIS proteins, the yeast protein stimulates RNA polymerase II to cleave the nascent RNA transcript and to read-through an intrinsic block to elongation. Investigation of the protein-protein contacts between TFIIS and RNA polymerase II indicated that the carboxyl-terminal domain of the largest subunit, subunit four, and subunit seven of the polymerase are not required for TFIIS to promote cleavage and read-through by the polymerase. In addition the carboxyl-terminal half of the yeast TFIIS protein is sufficient for both of these in vitro activities. This result is consistent with the previous results demonstrating the carboxyl-terminus of mouse TFIIS was sufficient to activate RNA polymerase in vitro.

Regulation of microRNAs

Regulation of microRNAs PDF Author: Helge Großhans
Publisher: Springer Science & Business Media
ISBN: 1441978232
Category : Science
Languages : en
Pages : 175

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Book Description
Given this pervasiveness and importance of miRNA-mediated gene regulation, it should come as little surprise that miRNAs themselves are also highly regulated. However, the recent explosion of knowledge on this topic has been remarkable, providing a primary motivation for publication of this book. As miRNAs are transcribed by RNA polymerase II, the enzyme that also generates mRNAs, it was perhaps not unexpected that miRNA transcription would be subject to regulation, and we have willfully mitted this aspect from this monograph. However, what has been unexpected is the extent of post-transcriptional regulation of miRNAs that is illustrated in this book.

Eukaryotic RNases and their Partners in RNA Degradation and Biogenesis

Eukaryotic RNases and their Partners in RNA Degradation and Biogenesis PDF Author:
Publisher: Academic Press
ISBN: 0124047459
Category : Science
Languages : en
Pages : 295

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Book Description
This special issue of The Enzymes is targeted towards researchers in biochemistry, molecular and cell biology, pharmacology, and cancer. This volume discusses Eukaryotic RNases and their partners in RNA degradation and biogenesis. - Contributions from leading authorities - Informs and updates on all the latest developments in the field