Multiple proteins bind to VA RNA genes of adenovirus type 2.

Dyke, Michael W. Van; Roeder, R G

doi:10.1128/mcb.7.3.1021

Cited by 36 publications

(23 citation statements)

References 45 publications

(47 reference statements)

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“…On the basis of further experiments carried out in vitro, it was suggested that NF1 can function as a general pol III transcription factor, influencing expression of a range of class III genes (31). The study, however, ignored an earlier paper from the same laboratory that had concluded that NF1 has no effect on VA gene transcription (45). We used ChIP assay to test whether NF1 is present at class III genes in vivo.…”

Section: Nf1 and Tfiia Are Not Detected At Pol Iii-transcribed Genes mentioning

confidence: 97%

Human La is found at RNA polymerase III-transcribed genes in vivo

Fairley

Kantidakis

Kenneth

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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The human La autoantigen can bind to nascent RNA transcripts and has also been postulated to act as an RNA polymerase III (pol III) transcription initiation and termination factor. Here, we show by chromatin immunoprecipitation (ChIP) that La is associated with pol III-transcribed genes in vivo. In contrast, the Ro autoantigen, which can also bind pol III transcripts, is not found at these genes. The putative pol III transcription factors NF1 and TFIIA are also not detected at class III genes. Binding of La remains when transcription is repressed at mitosis and does not correlate with the presence of polymerase at the gene. However, gene occupancy depends on the phosphorylation status of La, with the less prevalent, unphosphorylated form being found selectively on pol III templates.La ͉ NF1 ͉ TFIIA ͉ chromatin immunoprecipitation ͉ RNA polymerase III transcription R NA polymerase III (pol III) is responsible for the production of a number of short, untranslated RNAs that are essential for cellular growth; pol III transcription is therefore a major determinant of the biosynthetic capacity of cells (reviewed in refs. 1 and 2). The key to understanding how this influence is regulated is the identification of factors that can control expression. Pol III promoters can be divided into three types that utilize different transcription factors for their activity. Type 1 and 2 promoters, exemplified by the 5S rRNA and tRNA promoters, respectively, recruit TFIIIA and͞or TFIIIC, through binding to sequence-specific elements within the gene (reviewed in refs. 3 and 4). TFIIIC in turn recruits a TFIIIB complex containing Brf1, TBP, and Bdp1, which is able to recruit the polymerase through protein-protein interactions. In contrast, type 3 promoters, such as that for U6 small nuclear RNA (snRNA), contain gene external elements that are recognized by the SNAPc complex and TFIIIB (4); however, the TFIIIB in this case differs from that used for type 1 and type 2 promoters in utilizing Brf2 instead of Brf1 (4, 5). In addition to these basal transcription factors, a number of other proteins have been found to influence pol III transcription in mammals. Some of these regulators have been shown to associate with pol III templates in vivo, including c-Myc (6), RB (7), ␤-actin (8), and the protein kinase CK2 (9, 10). However, there are a number of other proteins, including La, TFIIA and NF1, for which a role in pol III transcription has been suggested from experiments in vitro, but which has not been confirmed in vivo.The human autoantigen La is a highly abundant protein found associated with newly synthesized pol III products (11,12). La binds to nascent pol III transcripts through the UUU-OH at their 3Ј ends, which corresponds to the pol III termination signal, and also has affinity for their 5Ј ends (13-15). One of the functions of La is in the stabilization of these transcripts, which promotes their processing to the mature forms (16). However, La has also been reported to have effects on pol III transcription. Immunodepletion of La...

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Section: Nf1 and Tfiia Are Not Detected At Pol Iii-transcribed Genes mentioning

confidence: 97%

Human La is found at RNA polymerase III-transcribed genes in vivo

Fairley

Kantidakis

Kenneth

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

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“…1). The deoxyribonuclease I (DNase I) footprinting pattern observed for the USF-TFIID-ML promoter complex (lane 3) analyzed by DNase I footprinting as described (10,16). The products of adenosine-(A) and guanosine-specific (G) chemical-sequencing reactions (24) were used as markers.…”

Section: G Ene Activity In Eukaryotes Ismentioning

confidence: 99%

Physical Analysis of Transcription Preinitiation Complex Assembly on a Class II Gene Promoter

Dyke

Roeder

Sawadogo

1988

Science

313

185

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Transcription of protein-encoding genes by human RNA polymerase II requires multiple ancillary proteins (transcription factors). Interactions between these proteins and the promoter DNA of a viral class II gene (the major late transcription unit of adenovirus) were investigated by enzymatic and chemical footprinting. The experiments indicated that the assembly of functionally active RNA polymerase II-containing transcription preinitiation complexes requires a complete set of transcription factors, and that both specific protein-DNA and protein-protein interactions are involved. This allows individual steps along the transcription reaction pathway to be tested directly, thus providing a basis for understanding basic transcription initiation mechanisms as well as the regulatory processes that act on them.

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“…DNase I cleavage was initiated by the addition of 3 ng of DNase I and allowed to proceed for 30 s at room temperature. Termination of the DNase cleavage reaction, DNA fragment purification, and gel electrophoresis followed standard protocols (24). Footprinting of other transcription complexes (i.e., preinitiation, energy dependent, and postinitiation) required a second incubation step, as described previously (25).…”

Section: Methodsmentioning

confidence: 99%

DNA-binding and transcriptional properties of human transcription factor TFIID after mild proteolysis.

Dyke¹,

Sawadogo²

1990

Mol. Cell. Biol.

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The existence of separable functions within the human class H general transcription factor TFIID was probed for differential sensitivity to mild proteolytic treatment. Independent of whether TFIID was bound to DNA or free in solution, partial digestion with either one of a variety of nonspecific endoproteases generated a proteaseresistant protein product that retained specific DNA recognition, as revealed by DNase I footprinting. However, in contrast to native TFIID, which interacts with the adenovirus major late (ML) promoter over a very broad DNA region, partially proteolyzed TFIID interacted with only a small region of the ML promoter immediately surrounding the TATA sequence. This novel footprint was very similar to that observed with the TATA factor purified from yeast cells. Partially proteolyzed human TFIID could form stable complexes that were resistant to challenge by exogenous templates. It could also nucleate the assembly of transcription complexes on the ML promoter with an efficiency comparable to that of native TFIID, yielding similar levels of transcription initiation. These results suggest a model in which the human TFHID protein is composed of at least two different regions or polypeptides: a protease-resistant "core," which by itself is sufficient for promoter recognition and basal transcriptional levels, and a protease-sensitive "tail," which interacts with downstream promoter regions and may be involved in regulatory processes.The promoter of protein-encoding (class II) genes is composed of multiple DNA elements. The minimum (or basal) promoter often contains a sequence referred to as the TATA box, whereas regulation is conferred by one or several upstream regulatory elements (for a review, see references 13 and 27). Recognition of the promoter by the general transcription factors leads to the formation of stable complexes that signal a gene for transcription by RNA polymerase II. Template challenge analyses have revealed that a particular transcription factor, designated TFIID, is the essential component of these stable complexes and that the TATA box sequence alone is sufficient for TFIID binding (4,26). Human TFIID is required for transcription of a number of cellular and viral genes in vitro (15). Binding of TFIID to the promoter DNA has been shown to be facilitated by an activity designated TFIIA (4, 6, 17), although an absolute TFIIA requirement for stable complex formation as well as for efficient transcription has not always been found (19,26 Recently, a protein was isolated from yeast which can substitute for human TFIID in a heterologous in vitro system reconstituted with the other general transcription factors and RNA polymerase II from HeLa cells (2, 3). This yeast TFIID is a single polypeptide of 27,000 daltons (10), which can form stable preinitiation complexes with various TATA boxes in the absence of any other transcription factor (3). DNase I footprinting has revealed a small (16 bp) region of interaction for yeast TFIID with the ML TATA box (2, 10), and the same nucleotides ...

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Multiple proteins bind to VA RNA genes of adenovirus type 2.

Cited by 36 publications

References 45 publications

Human La is found at RNA polymerase III-transcribed genes in vivo

Human La is found at RNA polymerase III-transcribed genes in vivo

Physical Analysis of Transcription Preinitiation Complex Assembly on a Class II Gene Promoter

DNA-binding and transcriptional properties of human transcription factor TFIID after mild proteolysis.

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