The U6 gene of Saccharomyces cerevisiae is transcribed by RNA polymerase C (III) in vivo and in vitro.

Moenne, Alejandra; Camier, Sylvie; Anderson, Gordon J.; Margottin, Florence; Beggs, Jean D.; Sentenac, André

doi:10.1002/j.1460-2075.1990.tb08105.x

Cited by 100 publications

(88 citation statements)

References 31 publications

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“…partially purified TFIB fraction (35). Figure 8 shows that the SNR6 gene could be specifically transcribed, like the tRNAGlU3 gene, in the presence of the FT250 fraction (that contains B" and TBP) supplemented by rTFIB70 ( Figure 8, lane 1).…”

Section: Resultsmentioning

confidence: 98%

Interactions between yeast TFIIIB components

Huet¹,

Conesa²,

Manaud³

et al. 1994

Nucl Acids Res

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Section: Resultsmentioning

confidence: 98%

Interactions between yeast TFIIIB components

Huet¹,

Conesa²,

Manaud³

et al. 1994

Nucl Acids Res

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“…To test whether TFIIIC influences termination at T 5 in different sequence contexts, some of the templates in which the T 5 terminator is weakened were modified so as to allow for their TFIIIC-independent transcription. To this end, the transcribed portions of L(CAA)LR2, L_3ЈN and SCR1⌬_3ЈL were fused to the 5Ј-flanking region of the S. cerevisiae SNR6 gene, which is known to support TATA box-mediated TFIIIB assembly and transcription initiation in the absence of TFIIIC (13,40). As shown in Fig.…”

Section: Distinguishing Between Weak and Strong T 5 Terminator Sequenmentioning

confidence: 99%

Sequence Context Effects on Oligo(dT) Termination Signal Recognition by Saccharomyces cerevisiae RNA Polymerase III

Braglia

Percudani

Dieci

2005

Journal of Biological Chemistry

102

112

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Eukaryotic RNA polymerase (Pol) III terminates transcription at short runs of T residues in the coding DNA strand. By genomic analysis, we found that T 5 and T 4 are the shortest Pol III termination signals in yeasts and mammals, respectively, and that, at variance with yeast, oligo(dT) terminators longer than T 5 are very rare in mammals. In Saccharomyces cerevisiae, the strength of T 5 as a terminator was found to be largely influenced by both the upstream and the downstream sequence context. In particular, the CT sequence, which is naturally present downstream of T 5 in the 3-flank of some tDNAs, was found to act as a terminator-weakening element that facilitates translocation by reducing Pol III pausing at T 5 . In contrast, tDNA transcription termination was highly efficient when T 5 was followed by an A or G residue. Surprisingly, however, when a terminationproficient T 5 signal was taken out from the tDNA context and placed downstream of a fragment of the SCR1 gene, its termination activity was compromised, both in vitro and in vivo. Even the T 6 sequence, acting as a strong terminator in tRNA gene contexts, was unexpectedly weak within the SNR52 transcription unit, where it naturally occurs. The observed sequence context effects reflect intrinsic recognition properties of Pol III, because they were still observed in a simplified in vitro transcription system only consisting of purified RNA polymerase and template DNA. Our findings strengthen the notion that termination signal recognition by Pol III is influenced in a complex way by the region surrounding the T cluster and suggest that read-through transcription beyond T clusters might play a significant role in the biogenesis of class III gene products.Eukaryotic RNA polymerase (Pol) 1 III is unique among DNAdependent RNA polymerases in recognizing a simple run of T residues on the coding strand as a termination signal, in the apparent absence of accessory factors. The minimal signal thought to be sufficient to provoke Pol III termination varies among different eukaryotes. T 4 suffices for termination by Xenopus and human Pol III (1, 2), T 4 /T 5 for Schizosaccharomyces pombe Pol III (3), and T 5 /T 6 for Saccharomyces cerevisiae Pol III (4). Pol III termination within T clusters is generally heterogeneous and progressive (5), and termination efficiency tends to increase with the length of the T run (4). Mechanistically, Pol III termination involves extensive pausing, dictated by the T cluster itself (5, 6). Even during elongation, the addition of three UMP residues in succession is particularly slow (5). Pol III pausing at T clusters sets cycles of hydrolytic RNA chain retraction (7). Both pausing and the associated hydrolytic RNA cleavage are affected by mutations in the C160, C128, and C11 subunits of yeast Pol III (7-9). In the case of C128 and C11, the same mutations have also been shown to affect the termination properties of Pol III, thus suggesting a functional interplay between hydrolytic RNA cleavage and transcription termination (9, 10). The t...

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“…Although TFIIIC appears to be essential for transcription of most class III genes (including those for tRNA, VA RNA, and 5S RNA), it has been reported not to be involved in the transcription of 7SK and U6 genes in vertebrate cell extracts (Waldschmidt et al 1991;Lagna et al 1994) and to be conditionally required for U6 genes in yeast (Brow and Guthrie 1990;Moenne et al 1990;Bumol et al 1993). Although the earlier vertebrate studies did not exclude the requirement for a form of TFIIIC2 that does not bind the 13 box (for review, see Lagna et al 19941, more definitive studies with antibodies to TFIIICa have eliminated a role for TFIIIC2 in 7SK and U6 transcription in cell extracts (Lagna et al 1994).…”

Section: Modification Of Basic Transcription Factors As a Global Regumentioning

confidence: 99%

Cloning and characterization of a TFIIIC2 subunit (TFIIIC beta) whose presence correlates with activation of RNA polymerase III-mediated transcription by adenovirus E1A expression and serum factors.

Sinn¹,

Wang²,

Kovelman³

et al. 1995

Genes Dev.

102

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TFIIIC2 is a general factor essential for transcription of 5S RNA, tRNA, and VA RNA genes by mammalian RNA polymerase III and consists of two forms designated THIIC2a and TFIIIC2b. TFIIIC2a and TFIIIC2b share common subunits of 220, 102, 90, and 63 kD but differ with respect to transcription activity and the presence of a presumptive 110-kD subunit in the active form (TFIIIC2a). Because both forms can bind the promoter directly, a selective role for the 110-kD subunit in the regulation of RNA polymerase III activity has been suggested. To investigate this possibility, we have cloned and expressed a cDNA encoding the l l0-kD subunit (TFIIICB). Immunoprecipitation studies with anti-TFIIICI3 antibodies have confirmed that TFIIIC[3 is a bona fide subunit present only in TFIIIC2a, that TFIIIC2a and the general factor TFIIIC1 are associated in unfractionated extracts, and that previously undetected polypeptides (potential TFIIIC1 subunits) can be isolated in association with TFIIIC2a. Previous studies have shown that increases in RNA polymerase III activity during infection of cells by adenovirus (with concomitant E1A expression) or during cell growth at high serum concentration results from an increased activity in the TFIIIC fraction. Studies with antibodies to TFIIICB have shown that this is strongly correlated with a selective increase in the cellular concentration of the TFIIIC~ l l0-kD subunit and a concomitant rise in the ratio of the active-to-inactive forms of TFIIIC2.

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The U6 gene of Saccharomyces cerevisiae is transcribed by RNA polymerase C (III) in vivo and in vitro.

Cited by 100 publications

References 31 publications

Interactions between yeast TFIIIB components

Interactions between yeast TFIIIB components

Sequence Context Effects on Oligo(dT) Termination Signal Recognition by Saccharomyces cerevisiae RNA Polymerase III

Cloning and characterization of a TFIIIC2 subunit (TFIIIC beta) whose presence correlates with activation of RNA polymerase III-mediated transcription by adenovirus E1A expression and serum factors.

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