Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound   Target Concepts: Gene/Protein Disease Symptom Drug Enzyme Compound

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Query: EC:2.7.7.6 (RNA polymerase)
34,946 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The phosphorylation, DNA-binding and dimerization properties of both forms of the RNA polymerase I transcription factor UBF were studied and compared. Tryptic peptide maps of in vivo 32P-labeled UBF contained four phospho-peptides. Two of these peptides are predicted to derive from the serine-rich, carboxyl-terminal of UBF. This region contains nine consensus phosphorylation sites for casein kinase II, and is one of the regions phosphorylated in vitro by casein kinase II. Analysis of the DNA-binding properties of recombinant forms of UBF1 and UBF2 by Southwestern blots revealed: (1) a role for the NH2-terminal 102 amino acid domain of UBF1/UBF2 in DNA-binding; (2) the importance of the bases from -106 to -101 of the rat ribosomal DNA promoter for the binding of UBF; and (3) functional differences between UBF1 and UBF2. Glutaraldehyde cross-linking and overlay assays using recombinant forms of UBF1 and UBF2 demonstrated that the molecules can form both homodimers and heterodimers. These assays also demonstrated that the NH2-terminal 102 amino acids of UBF plays a significant role in dimerization and that other domains contribute to dimerization. The dimerization properties of recombinant forms of UBF1 and UBF2 were different, suggesting that the HMG box 2 of UBF1, which is partially deleted in UBF2, also contributes to UBF dimerization.
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PMID:Analysis of the phosphorylation, DNA-binding and dimerization properties of the RNA polymerase I transcription factors UBF1 and UBF2. 156 Oct 86

We have analyzed the expression, phosphorylation, and localization of the ribosomal DNA transcription factors UBF1 and UBF2 in Chinese hamster ovary cells in response to serum deprivation. In vivo labeling experiments demonstrate that UBF1 and UBF2 are phosphoproteins. Phosphoamino acid analysis of the in vivo labeled proteins demonstrate that UBF is phosphorylated on serine residues. Following serum deprivation there is no alteration in the cellular levels of UBF1 and UBF2 as determined by Western blotting, but there is an 80% reduction in the level of phosphorylation of UBF compared with logarithmically growing cells. Following serum deprivation there is a redistribution of UBF between the nucleolus, the nucleus, and the cytoplasm. Phosphatase-treated UBF demonstrated a reduced ability to rescue transcription by RNA polymerase I from the rDNA spacer promoter in vitro. These findings suggest that phosphorylation of UBF is a prerequisite for transactivation of RNA polymerase I.
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PMID:Differential phosphorylation and localization of the transcription factor UBF in vivo in response to serum deprivation. In vitro dephosphorylation of UBF reduces its transactivation properties. 173 Jun

The human ribosomal RNA promoter contains two distinct control elements (UCE and core) both of which are recognized by the sequence-specific DNA binding protein UBF1, which has now been purified to apparent homogeneity. The purified factor activates RNA polymerase I (RNA pol I) transcription through direct interactions with either control element. A second RNA pol I transcription factor, designated SL1, participates in the promoter recognition process and is required to reconstitute transcription in vitro. Although SL1 alone has no sequence-specific DNA binding activity, deoxyribonuclease I footprinting experiments reveal that a cooperative interaction between UBF1 and SL1 leads to the formation of a new protein-DNA complex at the UCE and core elements. In vitro transcription experiments indicate that formation of the UBF1-SL1 complex is vital for transcriptional activation by UBF1. Thus, protein-protein interactions between UBF1 and SL1 are required for targeting of SL1 to cis-control sequences of the promoter.
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PMID:Functional cooperativity between transcription factors UBF1 and SL1 mediates human ribosomal RNA synthesis. 341 83

The human rRNA promoter contains two distinct cis-control sequences, the core and upstream control element (UCE), that serve as the target for binding cellular trans-activating proteins involved in transcription initiation by RNA polymerase I. One of these factors, SL1, has been extensively purified and shown to be a species-specific factor required to reconstitute in vitro RNA synthesis. DNAase footprinting revealed that although SL1 alone does not bind specifically to rRNA promoter sequences, a second factor, UBF1, recruits SL1 to the template and directs binding to an extended region encompassing sequences in the UCE. Analysis of mutant and human-mouse hybrid promoters indicate that protein-DNA interactions at the UCE modulate the efficiency of rRNA synthesis. Transcription from the human rRNA promoter appears to require an unusual set of protein-DNA transactions in which recognition and binding to an upstream cis-control element is carried out by one factor (UBF1), whereas activation requires an additional factor, SL1, acting in conjunction with UBF1 to trigger transcription.
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PMID:Human rRNA transcription is modulated by the coordinate binding of two factors to an upstream control element. 370 92

For efficient transcription from the rat ribosomal DNA (rDNA) promoter by RNA polymerase I in vitro, at least two transcription factors, rat UBF and rat SL-1, are required. Transcription cannot take place in vitro in the absence of SL-1. On the other hand, there is considerable difference of opinion concerning the necessity for UBF in in vitro transcription mediated by RNA polymerase 1, and the requirement for UBF is not clear. Mammalian cells code for UBF1 and UBF2, two forms of UBF that differ in HMG box-2, one of four HMG boxes or DNA-binding domains. We have used a monospecific antibody raised to recombinant rat UBF to determine whether UBF1 and UBF2 are required for RNA polymerase I-mediated transcription. This antibody can detect as little as 1.35 x 10(-15) moles of UBF1 or UBF2 in an immunoblot. Fractionated extracts that were competent for transcription had no detectable UBF1 or UBF2 when assayed in immunoblots with this antiserum. This evidence supports the hypothesis that UBF is not required for transcription of the rat rDNA promoter in vitro and most likely functions as an auxillary transcription factor. In addition, we have fractionated rat UBF1 from UBF2 and tested each of them in in vitro transcription assays in which the 45S or spacer rDNA promoter template is limiting. UBF1 can activate transcription from either the 45S or spacer promoter under these conditions, whereas UBF2 cannot. This implies that there is a functional difference in the transactivation of RNA polymerase I by UBF1 and UBF2 in vitro.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Transcription from the rat 45S ribosomal DNA promoter does not require the factor UBF. 801 25

rRNA synthesis decreases significantly during the differentiation of rat L6 myoblasts to myotubes. Nuclear run-on assays demonstrated that the decrease was attributable to decreased rates of rRNA gene transcription. Immunoblot analysis indicated a marked reduction in amounts of the RNA polymerase I transcription factors UBF1 and UBF2 (upstream binding factors 1 and 2, respectively). The levels of these factors dropped in parallel with the down-shift in rRNA gene transcription. The amount of UBF does not fall due to a general decrease in cellular protein, as myosin heavy-chain protein accumulates markedly during this same time. RNA blots of total RNA isolated from myoblasts and differentiating myotubes showed a decrease in the mRNA for UBF, at the same time the mRNA for myogenin was accumulating. The down-shift in UBF mRNA levels preceded the decrease in the protein levels for UBF. There have been reports that the acute response of the rRNA gene transcription system to physiological signals in many systems involves an RNA polymerase I-associated factor. However, our results imply that the regulation of rRNA gene DNA transcription in response to physiological processes, such as differentiation, may involve multiple regulatory pathways.
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PMID:Coordinated decreases in rRNA gene transcription factors and rRNA synthesis during muscle cell differentiation. 839 56

The RNA polymerase I transcription factor, UBF, belongs to a family of high-mobility-group DNA-binding proteins. Here, a human autoantibody reactive with the nucleolus organizer regions (NOR) was used to select cDNA clones encoding the hamster transcription factors, UBF1 and UBF2. Comparison at the nucleotide level showed a high degree of homology with other mammalian upstream binding factors (UBF) already identified. The deduced amino acid sequences are identical for both UBF1 and UBF2, except for a 37 amino acid insertion found in UBF1. This insertion is completely conserved among mammalian UBF1 which indicates a putative role of this region on the function of this transcript.
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PMID:Cloning and sequencing of the genes encoding the hamster ribosomal transcription factors UBF1 and UBF2. 891 62

Phytohaemagglutinin stimulates lymphoid cells to initiate active cell division which is tightly coordinated with transcription of ribosomal RNA genes. Nuclear Run-On assays demonstrated that treatment of peripheral blood lymphocytes with PHA (10 microg/ml) resulted in maximal rRNA synthesis after 64 hrs. In contrast, mRNA levels for upstream binding factor (UBF)1 and UBF2, as measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern blotting, increased relatively rapidly within 3 to 6 hrs. and remained elevated for at least the next 60 hrs. We further showed that exponentially growing cells of promyelocytic leukemia line HL-60 contained the same amounts of UBF1 and UBF2 mRNAs as phytohaemagglutinin (PHA)-stimulated lymphocytes for 6 hrs. Growth arrest of HL-60 cells, caused by 10 nM phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced monocytic/macrophage-like differentiation for 72 hrs., has been accompanied by a 50% decrease in UBF1/2 mRNAs expression. The lowest concentrations of UBF1/2 mRNAs were revealed in non-dividing terminally differentiated granulocytes. Regardless the activity of RNA polymerase I transcription and cell division rate, UBF1 mRNA levels prevailed over UBF2 mRNA levels in all human blood cell populations tested. Our results suggest that UBF gene expression is an important regulatory mechanism involved in the acceleration and possibly deceleration of rDNA transcription observed during mitogenic stimulation and inhibition of blood cells.
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PMID:Early gene expression of both RNA polymerase I transcription factors UBF1 and UBF2 precedes ribosomal RNA synthesis during lymphocyte mitogenic stimulation. 959 85

A novel RNA polymerase I (RPI) driven reporter gene has been used to investigate the in vivo role of the architectural ribosomal transcription factor UBF in gene activation and species specificity. It is shown that the level of UBF overexpression in NIH3T3 cells leads to a proportionate increase in the activities of both reporter and endogenous ribosomal genes. Further, co-expression of UBF antisense RNA suppresses reporter gene expression. Thus, UBF is limiting for ribosomal transcription in vivo and represents a potential endogenous ribosomal gene regulator. In contrast to some in vitro studies, in vivo, the mammalian and Xenopus forms of UBF1 show an equal ability to activate a mouse RPI promoter. This activity is severely impaired in mutants compromised for either dimerization or DNA binding. Similarly, the natural UBF2 splice variant shows a severely impaired capacity to activate RPI transcription. The data strongly suggest that UBF predominantly regulates ribosomal transcription by binding to and activating the ribosomal genes, but does not eliminate a possible secondary role in titrating ribosomal gene repressors such as Rb. Consistent with the DNA folding ability and cellular abundance of the UBF, we suggest that the protein may regulate a structural transition between the potentially active and active chromatin states.
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PMID:Cellular regulation of ribosomal DNA transcription:both rat and Xenopus UBF1 stimulate rDNA transcription in 3T3 fibroblasts. 992 57

beta-Catenin signaling plays an important role in the development of many organisms and has a key part in driving the malignant transformation of epithelial cells comprising a variety of cancers. beta-Catenin can activate gene expression through its association with transcription factors of the lymphoid enhancer factor 1 (LEF-1)/T-cell factor (TCF) family. We designed a screen in human cells to identify novel genes that activate a beta-catenin-LEF/TCF-responsive promoter and isolated the high-mobility group box transcription factor, UBF2. UBF1 and UBF2 are splice variants of a common precursor RNA. Although UBF1 has been shown to activate RNA polymerase I-regulated genes, the function of UBF2 has remained obscure. Here, we show for the first time that both UBF1 and UBF2 activate RNA polymerase II-regulated promoters. UBF2 associates with LEF-1, as shown by coimmunoprecipitation experiments, and potentiates transcriptional activation stimulated by LEF-1/beta-catenin from a synthetic promoter with multimerized LEF/TCF binding sites and a natural cyclin D1 promoter with consensus LEF/TCF binding sites. Downregulation of endogenous UBF expression using an RNA interference approach reduces transcriptional activation of a beta-catenin-LEF/TCF-responsive promoter by means of overexpressed beta-catenin, further implicating UBF as a transcriptional enhancer of the beta-catenin pathway.
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PMID:A functional screen in human cells identifies UBF2 as an RNA polymerase II transcription factor that enhances the beta-catenin signaling pathway. 1274 95


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