Transcription termination by RNA polymerase III: uncoupling of polymerase release from termination signal recognition (original) (raw)
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A model for transcription termination by RNA polymerase I
Cell, 1994
The transcription termination site for yeast RNA polymerase I requires not only an 11 bp binding site for Reb1p, but also about 46 bp of 5' flanking sequence. We propose that Reb1p bound to its site is part of a pause element, while the 5' flanking sequence contains a release element. Pausing requires little other than the DNA-binding domain of Reb1p and is not specific for polymerase I. The release element, however, can be polymerase specific. We propose a general model for eukaryotic transcription terminators in which termination occurs when a relatively nonspecific signal induces polymerase to pause in the context of a release element.
Transcription termination by the eukaryotic RNA polymerase III
Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms, 2013
RNA polymerase (pol) III transcribes a multitude of tRNA and 5S rRNA genes as well as other small RNA genes distributed through the genome. By being sequence-specific, precise and efficient, transcription termination by pol III not only defines the 3′ end of the nascent RNA which directs subsequent association with the stabilizing La protein, it also prevents transcription into downstream DNA and promotes efficient recycling. Each of the RNA polymerases appears to have evolved unique mechanisms to initiate the process of termination in response to different types of termination signals. However, in eukaryotes much less is known about the final stage of termination, destabilization of the elongation complex with release of the RNA and DNA from the polymerase active center. By comparison to pols I & II, pol III exhibits the most direct coupling of the initial and final stages of termination, both of which occur at a short oligo(dT) tract on the nontemplate strand (dA on the template) of the DNA. While pol III termination is autonomous involving the core subunits C2 and probably C1, it also involves subunits C11, C37 and C53, which act on the pol III catalytic center and exhibit homology to the pol II elongation factor TFIIS, and TFIIFα/β respectively. Here we compile knowledge of pol III termination and associate mutations that affect this process with structural elements of the polymerase that illustrate the importance of C53/37 both at its docking site on the pol III lobe and in the active center. The models suggest that some of these features may apply to the other eukaryotic pols.
Transcription termination by nuclear RNA polymerases
2009
Gene transcription in the cell nucleus is a complex and highly regulated process. Transcription in eukaryotes requires three distinct RNA polymerases, each of which employs its own mechanisms for initiation, elongation, and termination. Termination mechanisms vary considerably, ranging from relatively simple to exceptionally complex. In this review, we describe the present state of knowledge on how each of the three RNA polymerases terminates and how mechanisms are conserved, or vary, from yeast to human.
Molecular cell, 2015
Understanding the mechanism of transcription termination by a eukaryotic RNA polymerase (RNAP) has been limited by lack of a characterizable intermediate that reflects transition from an elongation complex to a true termination event. While other multisubunit RNAPs require multipartite cis-signals and/or ancillary factors to mediate pausing and release of the nascent transcript from the clutches of these enzymes, RNAP III does so with precision and efficiency on a simple oligo(dT) tract, independent of other cis-elements or trans-factors. We report an RNAP III pre-termination complex that reveals termination mechanisms controlled by sequence-specific elements in the non-template strand. Furthermore, the TFIIF-like RNAP III subunit C37 is required for this function of the non-template strand signal. The results reveal the RNAP III terminator as an information-rich control element. While the template strand promotes destabilization via a weak oligo(rU:dA) hybrid, the non-template stra...
Molecular Cell, 2002
berlin, 1992, Nudler et al., 1994; Zaychikov et al., 1995). The 3Ј terminal 8 nucleotides (nt) of the RNA are hydrogen bonded to the DNA template strand, forming the RNA:DNA hybrid inside the bubble (Zaychikov et al., 1995; Sidorenkov et al., 1998). The hybrid and approximately six more RNA residues lie inside RNAP (Komis-Frederick, Maryland 21702 sarova and Kashlev, 1998; Korzheva et al., 2000). It has 2 Goucher College been shown that the RNA:DNA hybrid is crucial for main-1021 Dulaney Valley Road taining the EC integrity (Korzheva et al., 1998; Sidoren-Baltimore, Maryland 21204 kov et al., 1998). Additionally, interaction between RNAP and single-stranded RNA upstream from the hybrid was suggested to stabilize the EC (von Hippel, 1998; Wilson Summary et al., 1999). Intrinsic terminators, which efficiently disrupt the EC, Passage of E. coli RNA polymerase through an intrinsic encode a stable stem-loop structure (hairpin) followed transcription terminator, which encodes an RNA hairby a U-rich sequence in the RNA (Yager and von Hippel,
Mechanism of Eukaryotic RNA Polymerase III Transcription Termination
Science, 2013
Gene expression in organisms involves many factors and is tightly controlled. Although much is known about the initial phase of transcription by RNA polymerase III (Pol III), the enzyme that synthesizes the majority of RNA molecules in eukaryotic cells, termination is poorly understood. Here, we show that the extensive structure of Pol III–synthesized transcripts dictates the release of elongation complexes at the end of genes. The poly-T termination signal, which does not cause termination in itself, causes catalytic inactivation and backtracking of Pol III, thus committing the enzyme to termination and transporting it to the nearest RNA secondary structure, which facilitates Pol III release. Similarity between termination mechanisms of Pol III and bacterial RNA polymerase suggests that hairpin-dependent termination may date back to the common ancestor of multisubunit RNA polymerases.
Distinguishing Core and Holoenzyme Mechanisms of Transcription Termination by RNA Polymerase III
Molecular and Cellular Biology, 2013
Transcription termination by RNA polymerase (Pol) III serves multiple purposes; it delimits interference with downstream genes, forms 3= oligo(U) binding sites for the posttranscriptional processing factor, La protein, and resets the polymerase complex for reinitiation. Although an interplay of several Pol III subunits is known to collectively control these activities, how they affect molecular function of the active center during termination is incompletely understood. We have approached this using immobilized Pol III-nucleic acid scaffolds to examine the two major components of termination, transcription pausing and RNA release. This allowed us to distinguish two mechanisms of termination by isolated Saccharomyces cerevisiae Pol III. A core mechanism can operate in the absence of C53/37 and C11 subunits but requires synthesis of 8 or more 3= U nucleotides, apparently reflecting inherent sensitivity to an oligo(rU•dA) hybrid that is the termination signal proper. The holoenzyme mechanism requires fewer U nucleotides but uses C53/37 and C11 to slow elongation and prevent terminator arrest. N-terminal truncation of C53 or point mutations that disable the cleavage activity of C11 impair their antiarrest activities. The data are consistent with a model in which C53, C37, and C11 activities are functionally integrated with the active center of Pol III during termination.
Molecular and cellular biology, 1996
We have studied the in vitro elongation and termination properties of several yeast RNA polymerase III (pol III) mutant enzymes that have altered in vivo termination behavior (S. A. Shaaban, B. M. Krupp, and B. D. Hall, Mol. Cell. Biol. 15:1467-1478, 1995). The pattern of completed-transcript release was also characterized for three of the mutant enzymes. The mutations studied occupy amino acid regions 300 to 325, 455 to 521, and 1061 to 1082 of the RET1 protein (P. James, S. Whelen, and B. D. Hall, J. Biol. Chem. 266:5616-5624, 1991), the second largest subunit of yeast RNA pol III. In general, mutant enzymes which have increased termination require a longer time to traverse a template gene than does wild-type pol III; the converse holds true for most decreased-termination mutants. One increased-termination mutant (K310T I324K) was faster and two reduced termination mutants (K512N and T455I E478K) were slower than the wild-type enzyme. In most cases, these changes in overall elonga...