The Saccharomyces cerevisiae SPT14 gene is essential for normal expression of the yeast transposon, Ty, as well as for expression of the HIS4 gene and several genes in the mating pathway

Fassler, Jan S.; Gray, William M.; Lee, Jean Pyo; Yu, Guoying Karen; Gingerich, Gregory

doi:10.1007/bf00290682

Cited by 29 publications

(22 citation statements)

References 49 publications

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“…From this analysis, m a n y of the known components of the holoenzyme can be accounted for, although a number of polypeptides remain unidentified. Some of these components may be encoded by genes that have been identified through other genetic screens (Fassler et al 1991;Berger et al 1992;Amakasu et al 1993;Pina et al 1993).…”

Section: A T a T G G C A G A G T T A A G C G T A T G T A T G T T A T mentioning

confidence: 99%

Association of an activator with an RNA polymerase II holoenzyme.

Hengartner¹,

Thompson²,

Zhang³

et al. 1995

Genes Dev.

222

200

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RNA polymerase II holoenzymes have been described that consist of RNA polymerase II, a subset of general transcription factors, and four SRB proteins. The SRB proteins, which were identified through a selection for genes involved in transcription initiation by RNA polymerase II in vivo, are a hallmark of the holoenzyme. We report here the isolation and characterization of additional SRB genes. We show that the products of all nine SRB genes identified thus far are components of the RNA polymerase II holoenzyme and are associated with a holoenzyme subcomplex termed the mediator of activation. The holoenzyme is capable of responding to a transcriptional activator, suggesting a model in which activators function, in part, through direct interactions with the holoenzyme. Immunoprecipitation experiments with anti-SRB5 antibodies demonstrate that the acidic activating domain of VPI6 specifically binds to the holoenzyme. Furthermore, the holoenzyme and the mediator subcomplex bind to a VP16 affinity column. These results provide a more complete description of the RNA polymerase II holoenzyme and suggest that this form of the transcription apparatus can be recruited to promoters via direct interactions with activators.[Key Words: RNA polymerase II; holoenzyme; carboxy-terminal domain; genetic suppressors; transcription initiation; SRBs; transcription activation] Received December 13, 1994; revised version accepted March 14, 1995.Large muhisubunit complexes containing RNA polymerase II, a subset of the general transcription factors, and additional factors implicated in regulation of transcription initiation in vivo, can assemble independently of promoter DNA (Kim et al. 1994;Koleske and Young 1994). These complexes, termed RNA polymerase II holoenzymes, have been purified from Saccharomyces cerevisiae. The larger form of holoenzyme contains RNA polymerase II, TFIIB, TFIIF, TFIIH, and SRB (suppressor of RNA polymerase B) proteins (Koleske and Young 1994). Another form of holoenzyme has been described that contains RNA polymerase II, TFIIF, and SRB proteins but lacks TFIIB and TFIIH (Kim et al. 1994). The two holoenzyme forms may exist simultaneously in vivo, or the isolation of the smaller complex may be a consequence of the instability of the RNA polymerase II holoenzyme during purification.Selective transcription initiation in vitro by the 12-subunit core RNA polymerase II was shown previously to require the action of at least five general initiation factors: TATA-binding protein (TBPJ, TFIIB, TFIIE, TFIIF, and TFIIH (for review, see Conaway and Conaway 1993;Zawel and Reinberg 1993). Consistent with these data, selective transcription initiation in vitro with the larger form of RNA polymerase II holoenzyme required TBP and TFIIE (Koleske and Young 1994), and initiation with the smaller form required TBP, TFIIB, TFIIE, and TFIIH (Kim et al. 1994).The holoenzymes were discovered by virtue of their association with SRB proteins. SRB genes were obtained through a genetic selection designed to identify genes involved in RNA ...

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Section: A T a T G G C A G A G T T A A G C G T A T G T A T G T T A T mentioning

confidence: 99%

Association of an activator with an RNA polymerase II holoenzyme.

Hengartner¹,

Thompson²,

Zhang³

et al. 1995

Genes Dev.

222

200

View full text Add to dashboard Cite

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“…Partial sequence analysis (Fig. 1B) revealed 99% identity over 613 base pairs to the SPT14gene [9] and its 3' flanking region (data not shown). As the restriction maps of CWH6 and SPT14…”

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confidence: 97%

“…SPT14 was initially identified as a suppressor of Ty insertion in the HIS4 promoter region and proposed to function as a trans-acting transcription factor [9]. However, the Sptl4-protein sequence showed no similarity to any known transcription factors.…”

mentioning

confidence: 99%

Identification of SPT14/CWH6 as the yeast homologue of hPIG-A, a gene involved in the biosynthesis of GPI anchors

Vossen

Ram

Klis

1995

Biochimica et Biophysica Acta (BBA) - General Subjects

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“…The effects of sptl3 mutations on a-specific genes have been attributed to reduced expression of the MATaJ gene, which is required for their expression. We have recently found that the MFA2 gene appears to be more sensitive to the absence of SPT13 than other previously measured a-specific genes are (15,17). The effect of sptl3 mutations on the expression of a-specific genes cannot be attributed to reductions in MATo expression.…”

Section: Discussionmentioning

confidence: 76%

SPT13 (GAL11) of Saccharomyces cerevisiae negatively regulates activity of the MCM1 transcription factor in Ty1 elements.

Yu¹,

Fassler²

1993

Mol. Cell. Biol.

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The Ty transposable elements of Saccharomyces cerevisiae consist of a single large transcription unit whose expression is controlled by a combination of upstream and downstream regulatory sequences. Errede (B. Errede, Mol. Cell. Biol. 13:57-62, 1993) has shown that among the downstream control sequences is a binding site for the transcription factor, MCM1. A small restriction fragment containing the Tyl MCMI-binding site exhibits very weak activation of heterologous gene expression. The absence of SPT13 (GAL1l) causes a dramatic increase in activity directed by these sequences. This effect is mediated through the MCM1-binding site itself. MCM1 mRNA and protein levels, as well as its affinity for its binding site, are unchanged in the absence of SPT13. Our results suggest that SPT13 has a role in the negative control of MCM1 activity that is likely to be posttranslational. A role for SPT13 in the negative regulation of the activity of the Tyl MCMI-binding site is consistent with our previous proposal that sptl3-mediated suppression of Ty insertion mutations could be attributed to the loss of negative regulation of genes adjacent to Ty elements.The Ty (transposable) elements of Saccharomyces cerevisiae are a family of retrovirus-like transposons consisting of a unique 5.3-kb (e) region flanked by 330-bp direct repeats (8). Transcription of these elements initiates in one 8 repeat and terminates in the other (9). In addition to the expected upstream regulatory elements located within 8 sequences (9, 29), extensive molecular analysis has revealed additional regulatory elements located inside the transcription unit (13, 37). Several downstream regulatory elements in Tyl elements that function independently as weak activators of a heterologous reporter gene have been identified and characterized (5-7). One element is located just downstream of the Ty promoter and consists of a STE12-binding site (5). Two other regulatory elements, block I and block II, are located several hundred nucleotides downstream of the Ty transcriptional start site and contain homology to three different regulatory motifs including the simian virus 40 (SV40) enhancer core, the al-a2 diploid control signal, and the MCM1 recognition sequence or P site (7,11,13,14). The role of these sequences in promoting transcription of Ty elements is unknown. A 28-bp sequence containing block II is weakly capable of activating adjacent-gene transcription (7). Like expression of the Ty element itself, activation directed by these sequences is cell type specific and is not observed in diploid cells (7).Previous analysis of block II and adjacent sequences revealed the presence of two protein-binding sites (7). The binding site of one complex falls within block II; the second site maps immediately downstream of block II sequences (7). Errede has identified the block II-binding protein as the constitutive factor, MCM1 (11).We have been analyzing the function of the Ty regulatory elements by the isolation of a series of mutants altered in the expression of Ty-adja...

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The Saccharomyces cerevisiae SPT14 gene is essential for normal expression of the yeast transposon, Ty, as well as for expression of the HIS4 gene and several genes in the mating pathway

Cited by 29 publications

References 49 publications

Association of an activator with an RNA polymerase II holoenzyme.

Association of an activator with an RNA polymerase II holoenzyme.

Identification of SPT14/CWH6 as the yeast homologue of hPIG-A, a gene involved in the biosynthesis of GPI anchors

SPT13 (GAL11) of Saccharomyces cerevisiae negatively regulates activity of the MCM1 transcription factor in Ty1 elements.

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