An RNA Polymerase II Elongation Factor Encoded by the Human ELL Gene

Abstract The Ras signaling pathway in Saccharomyces cerevisiae controls cell growth via the cAMP-dependent protein kinase, PKA. Recent work has indicated that these effects on growth are due, in part, to the regulation of activities associated with the C-terminal domain (CTD) of the largest subunit of RNA polymerase II. However, the precise target of these Ras effects has remained unknown. This study suggests that Ras/PKA activity regulates the elongation step of the RNA polymerase II transcription process. Several lines of evidence indicate that Spt5p in the Spt4p/Spt5p elongation factor is the likely target of this control. First, the growth of spt4 and spt5 mutants was found to be very sensitive to changes in Ras/PKA signaling activity. Second, mutants with elevated levels of Ras activity shared a number of specific phenotypes with spt5 mutants and vice versa. Finally, Spt5p was efficiently phosphorylated by PKA in vitro. Altogether, the data suggest that the Ras/PKA pathway might be directly targeting a component of the elongating polymerase complex and that this regulation is important for the normal control of yeast cell growth. These data point out the interesting possibility that signal transduction pathways might directly influence the elongation step of RNA polymerase II transcription.

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Identification of rpo30, a vaccinia virus RNA polymerase gene with structural similarity to a eucaryotic transcription elongation factor

Molecular and Cellular Biology ◽

10.1128/mcb.10.10.5433-5441.1990 ◽

1990 ◽

Vol 10 (10) ◽

pp. 5433-5441

Author(s):

B Y Ahn ◽

P D Gershon ◽

E V Jones ◽

B Moss

Keyword(s):

Rna Polymerase ◽

Vaccinia Virus ◽

Rna Polymerase Ii ◽

Elongation Factor ◽

Viral Rna ◽

In Vitro Translation ◽

Virus Protein ◽

Transcriptional Elongation

Eucaryotic transcription factors that stimulate RNA polymerase II by increasing the efficiency of elongation of specifically or randomly initiated RNA chains have been isolated and characterized. We have identified a 30-kilodalton (kDa) vaccinia virus-encoded protein with apparent homology to SII, a 34-kDa mammalian transcriptional elongation factor. In addition to amino acid sequence similarities, both proteins contain C-terminal putative zinc finger domains. Identification of the gene, rpo30, encoding the vaccinia virus protein was achieved by using antibody to the purified viral RNA polymerase for immunoprecipitation of the in vitro translation products of in vivo-synthesized early mRNA selected by hybridization to cloned DNA fragments of the viral genome. Western immunoblot analysis using antiserum made to the vaccinia rpo30 protein expressed in bacteria indicated that the 30-kDa protein remains associated with highly purified viral RNA polymerase. Thus, the vaccinia virus protein, unlike its eucaryotic homolog, is an integral RNA polymerase subunit rather than a readily separable transcription factor. Further studies showed that the expression of rpo30 is regulated by dual early and later promoters.

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Negative Elongation Factor Is Required for the Maintenance of Proviral Latency but Does Not Induce Promoter-Proximal Pausing of RNA Polymerase II on the HIV Long Terminal Repeat

Molecular and Cellular Biology ◽

10.1128/mcb.01013-13 ◽

2014 ◽

Vol 34 (11) ◽

pp. 1911-1928 ◽

Cited By ~ 59

Author(s):

J. K. Jadlowsky ◽

J. Y. Wong ◽

A. C. Graham ◽

C. Dobrowolski ◽

R. L. Devor ◽

...

Keyword(s):

Rna Polymerase ◽

Long Terminal Repeat ◽

Rna Polymerase Ii ◽

Elongation Factor ◽

Terminal Repeat ◽

Negative Elongation Factor ◽

Proviral Latency

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Acute perturbation strategies in interrogating RNA polymerase II elongation factor function in gene expression

Genes & Development ◽

10.1101/gad.346106.120 ◽

2021 ◽

Vol 35 (3-4) ◽

pp. 273-285

Author(s):

Bin Zheng ◽

Yuki Aoi ◽

Avani P. Shah ◽

Marta Iwanaszko ◽

Siddhartha Das ◽

...

Keyword(s):

Gene Expression ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Elongation Factor ◽

Factor Function

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Crystal structure of the human transcription elongation factor DSIF hSpt4 subunit in complex with the hSpt5 dimerization interface

Biochemical Journal ◽

10.1042/bj20091422 ◽

2009 ◽

Vol 425 (2) ◽

pp. 373-380 ◽

Cited By ~ 22

Author(s):

Sabine Wenzel ◽

Berta M. Martins ◽

Paul Rösch ◽

Birgitta M. Wöhrl

Keyword(s):

Crystal Structure ◽

Escherichia Coli ◽

Transcription Factor ◽

Rna Polymerase ◽

Rna Polymerase Ii ◽

Transcriptional Activation ◽

Transcription Elongation ◽

Elongation Factor ◽

Structural Similarity ◽

Transcription Elongation Factor

The eukaryotic transcription elongation factor DSIF [DRB (5,6-dichloro-1-β-D-ribofuranosylbenzimidazole) sensitivity-inducing factor] is composed of two subunits, hSpt4 and hSpt5, which are homologous to the yeast factors Spt4 and Spt5. DSIF is involved in regulating the processivity of RNA polymerase II and plays an essential role in transcriptional activation of eukaryotes. At several eukaryotic promoters, DSIF, together with NELF (negative elongation factor), leads to promoter-proximal pausing of RNA polymerase II. In the present paper we describe the crystal structure of hSpt4 in complex with the dimerization region of hSpt5 (amino acids 176–273) at a resolution of 1.55 Å (1 Å=0.1 nm). The heterodimer shows high structural similarity to its homologue from Saccharomyces cerevisiae. Furthermore, hSpt5-NGN is structurally similar to the NTD (N-terminal domain) of the bacterial transcription factor NusG. A homologue for hSpt4 has not yet been found in bacteria. However, the archaeal transcription factor RpoE” appears to be distantly related. Although a comparison of the NusG-NTD of Escherichia coli with hSpt5 revealed a similarity of the three-dimensional structures, interaction of E. coli NusG-NTD with hSpt4 could not be observed by NMR titration experiments. A conserved glutamate residue, which was shown to be crucial for dimerization in yeast, is also involved in the human heterodimer, but is substituted for a glutamine residue in Escherichia coli NusG. However, exchanging the glutamine for glutamate proved not to be sufficient to induce hSpt4 binding.

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Heterogeneous nuclear ribonucleoprotein L-like (hnRNPLL) and elongation factor, RNA polymerase II, 2 (ELL2) are regulators of mRNA processing in plasma cells

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1214414109 ◽

2012 ◽

Vol 109 (40) ◽

pp. 16252-16257 ◽

Cited By ~ 23

Author(s):

M. J. Benson ◽

T. Aijo ◽

X. Chang ◽

J. Gagnon ◽

U. J. Pape ◽

...

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Plasma Cells ◽

Elongation Factor ◽

Mrna Processing ◽

Heterogeneous Nuclear Ribonucleoprotein ◽

Nuclear Ribonucleoprotein

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The FACT chromatin modulator: genetic and structure/function relationships

Biochemistry and Cell Biology ◽

10.1139/o04-050 ◽

2004 ◽

Vol 82 (4) ◽

pp. 419-427 ◽

Cited By ~ 23

Author(s):

Richard A Singer ◽

Gerald C Johnston

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Protein Unfolding ◽

Elongation Factor ◽

Yeast Genetics ◽

Genetic Studies ◽

Hmg Box ◽

Transcription Elongation Factor ◽

C Terminus ◽

Chromatin Configuration

The chromatin configuration of DNA inhibits access by enzymes such as RNA polymerase II. This inhibition is alleviated by FACT, a conserved transcription elongation factor that has been found to reconfigure nucleosomes to allow transit along the DNA by RNA polymerase II, thus facilitating transcription. FACT also reorganizes nucleosomes after the passage of RNA polymerase II, as indicated by the effects of certain FACT mutations. The larger of the two subunits of FACT is Spt16/Cdc68, while the smaller is termed SSRP1 (vertebrates) or Pob3 (budding yeast). The HMG-box domain at the C terminus of SSRP1 is absent from Pob3; the function of this domain for yeast FACT is supplied by the small HMG-box protein Nhp6. In yeast, this "detachable" HMG domain is a general chromatin component, unlike FACT, which is found only in transcribed regions and associated with RNA polymerase II. The several domains of the larger FACT subunit are also likely to have different functions. Genetic studies suggest that FACT mediates nucleosome reorganization along several pathways, and reinforce the notion that protein unfolding and (or) refolding is involved in FACT activity for transcription.Key words: nucleosomes, transcription, FACT, yeast, genetics.

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Genetic interaction between transcription elongation factor TFIIS and RNA polymerase II

Molecular and Cellular Biology ◽

10.1128/mcb.12.9.4142-4152.1992 ◽

1992 ◽

Vol 12 (9) ◽

pp. 4142-4152

Author(s):

J Archambault ◽

F Lacroute ◽

A Ruet ◽

J D Friesen

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Structural Integrity ◽

Genetic Interaction ◽

Elongation Factor ◽

Chemical Mutagenesis ◽

Yeast Genome ◽

Transcript Elongation

Little is known about the regions of RNA polymerase II (RNAPII) that are involved in the process of transcript elongation and interaction with elongation factors. One elongation factor, TFIIS, stimulates transcript elongation by binding to RNAPII and facilitating its passage through intrinsic pausing sites in vitro. In Saccharomyces cerevisiae, TFIIS is encoded by the PPR2 gene. Deletion of PPR2 from the yeast genome is not lethal but renders cells sensitive to the uracil analog 6-azauracil (6AU). Here, we show that mutations conferring 6AU sensitivity can also be isolated in the gene encoding the largest subunit of S. cerevisiae RNAPII (RPO21). A screen for mutations in RPO21 that confer 6AU sensitivity identified seven mutations that had been generated by either linker-insertion or random chemical mutagenesis. All seven mutational alterations are clustered within one region of the largest subunit that is conserved among eukaryotic RNAPII. The finding that six of the seven rpo21 mutants failed to grow at elevated temperature underscores the importance of this region for the functional and/or structural integrity of RNAPII. We found that the 6AU sensitivity of the rpo21 mutants can be suppressed by increasing the dosage of the wild-type PPR2 gene, presumably as a result of overexpression of TFIIS. These results are consistent with the proposal that in the rpo21 mutants, the formation of the RNAPII-TFIIS complex is rate limiting for the passage of the mutant enzyme through pausing sites. In addition to implicating a region of the largest subunit of RNAPII in the process of transcript elongation, our observations provide in vivo evidence that TFIIS is involved in transcription by RNAPII.

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The telomeric Cdc13–Stn1–Ten1 complex regulates RNA polymerase II transcription

Nucleic Acids Research ◽

10.1093/nar/gkz279 ◽

2019 ◽

Vol 47 (12) ◽

pp. 6250-6268 ◽

Cited By ~ 2

Author(s):

Olga Calvo ◽

Nathalie Grandin ◽

Antonio Jordán-Pla ◽

Esperanza Miñambres ◽

Noelia González-Polo ◽

...

Keyword(s):

Rna Polymerase ◽

Rna Polymerase Ii ◽

Genome Stability ◽

Elongation Factor ◽

Yeast Saccharomyces Cerevisiae ◽

Replication Fork Progression ◽

Genome Wide ◽

Length Regulation ◽

Transcription Regulators ◽

Rna Polymerase Ii Transcription

Abstract Specialized telomeric proteins have an essential role in maintaining genome stability through chromosome end protection and telomere length regulation. In the yeast Saccharomyces cerevisiae, the evolutionary conserved CST complex, composed of the Cdc13, Stn1 and Ten1 proteins, largely contributes to these functions. Here, we report genetic interactions between TEN1 and several genes coding for transcription regulators. Molecular assays confirmed this novel function of Ten1 and further established that it regulates the occupancies of RNA polymerase II and the Spt5 elongation factor within transcribed genes. Since Ten1, but also Cdc13 and Stn1, were found to physically associate with Spt5, we propose that Spt5 represents the target of CST in transcription regulation. Moreover, CST physically associates with Hmo1, previously shown to mediate the architecture of S-phase transcribed genes. The fact that, genome-wide, the promoters of genes down-regulated in the ten1-31 mutant are prefentially bound by Hmo1, leads us to propose a potential role for CST in synchronizing transcription with replication fork progression following head-on collisions.

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