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• W2016070620 abstract "Article15 September 1997free access Three classes of mammalian transcription activation domain stimulate transcription in Schizosaccharomyces pombe Jacques E. Remacle Corresponding Author Jacques E. Remacle Department of Cell Growth, Differentiation and Development (VIB-07), Flanders Interuniversity Institute for Biotechnology, Belgium Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Gerd Albrecht Gerd Albrecht Institute of Microbiology, Georg-August University, Grisebachstrasse 8, D-37077 Göttingen, Germany Search for more papers by this author Reginald Brys Reginald Brys Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Gerhard H. Braus Gerhard H. Braus Institute of Microbiology, Georg-August University, Grisebachstrasse 8, D-37077 Göttingen, Germany Search for more papers by this author Danny Huylebroeck Danny Huylebroeck Department of Cell Growth, Differentiation and Development (VIB-07), Flanders Interuniversity Institute for Biotechnology, Belgium Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Jacques E. Remacle Corresponding Author Jacques E. Remacle Department of Cell Growth, Differentiation and Development (VIB-07), Flanders Interuniversity Institute for Biotechnology, Belgium Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Gerd Albrecht Gerd Albrecht Institute of Microbiology, Georg-August University, Grisebachstrasse 8, D-37077 Göttingen, Germany Search for more papers by this author Reginald Brys Reginald Brys Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Gerhard H. Braus Gerhard H. Braus Institute of Microbiology, Georg-August University, Grisebachstrasse 8, D-37077 Göttingen, Germany Search for more papers by this author Danny Huylebroeck Danny Huylebroeck Department of Cell Growth, Differentiation and Development (VIB-07), Flanders Interuniversity Institute for Biotechnology, Belgium Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium Search for more papers by this author Author Information Jacques E. Remacle 1,2, Gerd Albrecht3, Reginald Brys2, Gerhard H. Braus3 and Danny Huylebroeck1,2 1Department of Cell Growth, Differentiation and Development (VIB-07), Flanders Interuniversity Institute for Biotechnology, Belgium 2Laboratory of Molecular Biology (CELGEN), University of Leuven, Herestraat 49, B-3000 Leuven, Belgium 3Institute of Microbiology, Georg-August University, Grisebachstrasse 8, D-37077 Göttingen, Germany *Corresponding author. E-mail: [email protected] The EMBO Journal (1997)16:5722-5729https://doi.org/10.1093/emboj/16.18.5722 PDFDownload PDF of article text and main figures. ToolsAdd to favoritesDownload CitationsTrack CitationsPermissions ShareFacebookTwitterLinked InMendeleyWechatReddit Figures & Info Representatives of three distinct classes of mammalian protein domain activating RNA polymerase II were fused to the yeast GAL4p DNA-binding domain. The resulting fusion proteins were tested in the fission yeast Schizosaccharomyces pombe for their ability to activate transcription of different reporter constructs containing GAL4-binding sites in positions close to or far from the TATA box. The acidic-rich activation domain of VP16 stimulates transcription in S.pombe from proximal and distal positions, suggesting that the mechanism of activation is conserved from man to budding and fission yeasts. Unlike in Saccharomyces cerevisiae, the glutamine-rich activation domains of Sp1, Oct1 and Oct2 activate transcription in S.pombe when tested in a proximal TATA box context. Similarly to mammalian cells, these domains are inactive or weakly active when tested in a distal position. Moreover, the proline-rich activation domains of AP-2 and CTF/NF1 display strong transcriptional activities from a TATA box-proximal position, and weak activities when tested in a remote position. Consequently, proline-rich and glutamine-rich activation domains act differently in S.cerevisiae and mammalian cells, but similarly in S.pombe and mammalian cells. Introduction Initiation of transcription in eukaryotes occurs through a complex set of DNA–protein and protein–protein interactions involving RNA polymerase II (which is presumably part of a complex holoenzyme), promoter DNA, a set of general transcription factors (GTFs) and adaptors, and promoter-specific regulatory proteins (Zawel and Reinberg, 1995; Künzler et al., 1996). The latter are sequence-specific DNA-binding proteins that in many cases have been shown to function as transcriptional activators. Typically, these activators are composed of distinct domains including separable DNA-binding and activation domains (Hope and Struhl, 1986; Keegan et al., 1986; for a review, see Triezenberg, 1995). The activation domains have been classified somewhat arbitrarily depending on whether they are rich in acidic amino acids, glutamine or proline. However, for the acidic-rich activation domains, recent observations revealed that specific patterns of hydrophobic and aromatic amino acids were equally or even more critical than the acidic ones (Cress and Triezenberg, 1991; Regier et al., 1993; Drysdale et al., 1995). These classes of activation domain differ significantly in their ability to activate transcription from distinct promoter positions in mammalian cells (Seipel et al., 1992). ‘Proximal’ activation domains, exemplified by glutamine-rich domains of Sp1, Oct1 and Oct2, stimulate transcription only from a position close to the TATA box, usually in response to a remote enhancer. Acidic-type activation domains, represented by yeast GAL4p and herpes simplex virus VP16, stimulate transcription from remote (e.g. enhancer) as well as proximal promoter positions. Finally, the proline-rich activation domains of AP-2 and CTF/NF1 display considerable proximal and low but significant remote promoter activity. Several mammalian proteins containing acidic-rich and/or proline-rich activation domains have been shown to stimulate transcription in the budding yeast (Lech et al., 1988; Metzger et al., 1988; Sadowski et al., 1988; Schena and Yamamoto, 1988; Struhl, 1988). Notably, the yeast transcription factor GAL4p, which has an acidic-type activation domain, also activates transcription in plant and mammalian cells, suggesting that its mechanism of activation is conserved (Ma et al., 1988; Webster et al., 1988). In contrast, the human Sp1 transcription factor, which contains well characterized glutamine-rich activation domains, fails to stimulate transcription in Saccharomyces cerevisiae even in concert with human TATA-binding protein (TBP) or human–yeast TBP hybrids (Ponticelli et al., 1995). The glutamine-rich activation domains of Oct1, Oct2 and Sp1, when fused to the DNA-binding domain of GAL4p, cannot stimulate transcription in S.cerevisiae from promoters containing GAL4-binding sites (Künzler et al., 1994). Consequently, in contrast to the acidic activation domains, the function of glutamine-rich domains in transcription differs between humans and S.cerevisiae. One explanation for this may be that glutamine-rich activation domains require for their function in mammals additional factors which are absent or have other functions in S.cerevisiae. For several aspects, such as chromosome size, centromere structure, intron splicing mechanism, cell cycle control and heat shock response, the fission yeast seems to be more closely related to mammalian cells than to budding yeast (Glick, 1996). In addition, as in mammalian cells, initiation of transcription from Schizosaccharomyces pombe promoters occurs ∼30 bp downstream of the TATA box, whereas in S.cerevisiae distances can vary between 40 and 120 bp (Li et al., 1994). Moreover, initiation from several mammalian promoters introduced into S.pombe occurs at the same site(s) as in mammalian cells (Toyama and Okayama, 1990). Since TFIIB and RNA polymerase II are mainly responsible for selecting the start site of transcription, this suggests a higher degree of conservation of the functions of these proteins between S.pombe and mammalian cells, when compared with S.cerevisiae. Several transcription factors have been cloned from S.pombe by other groups (Sugimoto et al., 1991; Tang et al., 1994; Wu and McLeod, 1995; Wilkinson et al., 1996). However, the characterization of their potential transcriptional activation domains (TADs) has not been performed yet. Therefore, there are hardly any data on the function of such domains at various promoter positions in S.pombe. Consequently, it is interesting to document whether mammalian transcriptional activators work in S.pombe as they do in mammalian cells or as they do in S.cerevisiae. We tested, in a systematic approach, representatives of the three classes of mammalian TAD in S.pombe. For this study, we used exactly the same GAL4DBD–TAD fusions that were tested in mammals and in S.cerevisiae (Seipel et al., 1992; Künzler et al., 1994). These hybrid proteins are based on the GAL4p DNA-binding domain (GAL4DBD; amino acids 1–93) fused to the activation domains from well-characterized mammalian transcription factors (Seipel et al., 1992; Künzler et al., 1994). These different GAL4DBD–TAD fusions were tested for transactivation of several S.pombe reporter constructs containing GAL4p-binding sites located at different positions from the TATA box. Acidic-type activation domains stimulate transcription strongly from positions both close to and remote from the TATA box. As in mammalian cells, proline-rich domains stimulate transcription strongly from a position close to the TATA box but only weakly from a remote position. Most importantly, glutamine-rich activation domains stimulate transcription in S.pombe, but only from a position close to the TATA box, just as is observed in mammals. These results indicate that, in contrast to S.cerevisiae, S.pombe is able to use glutamine-rich activation domains. These observations on the use of different types of activation domains in S.pombe significantly support the emerging picture that S.pombe, in many respects and in particular with regard to transcription, is much more related to and comparable with mammalian cells than S.cerevisiae. Results Overexpression of mammalian activation domains is toxic in the fission yeast TADs of different well-characterized mammalian transcription factors fused to the DNA-binding domain of GAL4p (GAL4DBD) have been analysed previously for transactivation in the budding yeast S.cerevisiae and in mammalian cells (Seipel et al., 1992; Künzler et al., 1994). Therefore, to allow comparison of the transcriptional activity of the different types of activation domains in S.pombe, S.cerevisiae and mammals, we tested in S.pombe exactly the same GAL4DBD–TADs fusions that were studied both in S.cerevisiae and in mammalian cells. We selected representatives of all three classes of activation domain from this collection of chimeric activators and used these in different S.pombe reporter strains (Table I and Figure 1). Acidic-rich activation domains are exemplified by different portions of the VP16 activation domain. Different portions of the AP-2 activation domain and the activation domain of CTF/NF1 represent the proline-rich class of activation domain. Finally, the class of glutamine-rich activation domains is represented by Oct1 and Oct2, and by different portions of the Sp1 activation domain. As a negative control, GAL4DBD was used without an activation domain. Chimeric activators were cloned into an episomal plasmid under the control of the thiamine-repressible S.pombe PHO4 promoter (Silvestre and Jacobs, 1997). Transcription from this promoter is induced on medium lacking thiamine. Figure 1.The arrangement of the GAL4p-binding sites in the different ADHmin reporter genes. The GAL4DBD–TAD fusion proteins were tested for transactivation in S.pombe by introducing the different expression plasmids in four S.pombe reporter strains: A1, A2, B and C. The construction of these strains is described in Materials and methods. The strain C (neo reporter gene) enables us to test the different GAL4DBD–TAD in a position remote (‘235 bp’) from the TATA box of the ADHmin promoter. For this, 5GAL4-binding sites were separated from the TATA box by three copies of a transcriptionally silent 71 bp DNA fragment (symbolized by the double arrow) of the firefly luciferase gene. For transactivation in a proximal position from the TATA box, we used strains A1 and A2, which contain a 17mer GAL4-binding site positioned immediately upstream of the ADH TATA box. The neo gene (strain A1) and the lacZ gene (strain A2) were used as reporter genes. The strain B (lacZ reporter gene) allows transactivation to be tested for in an intermediate position (‘87 bp’) upstream of the TATA box. To construct this strain, one copy of the same 71 bp luciferase DNA fragment was inserted between one GAL4-binding site and the ADH TATA box. Download figure Download PowerPoint Table 1. Summary of the transactivation results GAL4 fusion constructs Prevalent amino acid Net charge Activation in S.pombe (reporter constructs: Figure 1) TATA-proximal (A) 87 bp spacing (B) Remote (C) GAL4(1–93) − − − GAL4(1–93)–Sp1(132–243)1 Q 0 + +/− − GAL4(1–93)–Sp1(340–485)2 Q +1 + +/− − GAL4(1–93)–Oct1(175–269) Q +1 + +/− − GAL4(1–93)–Oct2(99–161) Q +2 ++ + +/− GAL4(1–93)–AP2(31–117)1 P>S + T +5 +++ +++ +/− GAL4(1–93)–AP2(31–76)2 P>S + T 0 ++ + +/− GAL4(1–93)–CTF(399–499) P>S + T +3 ++ ++ +/− GAL4(1–93)–VP16(c)(413–454) D + E −7 ++ ++ ++ GAL4(1–93)–VP16(413–490) D + E −18 +++ +++ +++ Amino acid positions are given in parentheses. The exact amino acid sequences of the fusion proteins are described elsewhere (Seipel et al., 1992). +++ indicates very strong, ++ strong, + medium, +/− weak and − no activation. 1 and 2 denote different TADs in the factor (Sp1) or different forms of the same factor (AP-2). All the different GAL4DBD–TAD expression plasmids were transformed into the S.pombe P2 strain. For each plasmid, one half of the transformation mixture was spread on selective medium containing thiamine (MMRT) and, therefore, expression of chimeric activators was repressed; the other half was plated on selective medium lacking thiamine (MMR), which enables synthesis of chimeric activator polypeptides. No differences in number and size of the colonies on both selection agar plates were detected with the GAL4DBD control plasmid (data not shown). When the GAL4DBD–TAD fusion genes were repressed, the URA4 colonies obtained on MMRT plates displayed the same size as the GAL4DBD control. However, when the fusion gene expressions were induced on medium lacking thiamine (MMR), no colony was obtained for the GAL4 hybrids with the proline-rich and acidic-rich activation domains. With the glutamine-rich activation domains, the size of these colonies was significantly smaller than the size of the colonies obtained with the GAL4DBD plasmid. To investigate this growth phenotype further, we measured the growth rate of yeast strains containing expression plasmids for the different types of activation domain in liquid culture. The cultures were cultivated in thiamine-containing medium (MMRT) and shifted to medium lacking thiamine (MMR). The growth rate (measured 35 h after the medium shift) of the strain expressing the GAL4DBD was 2.8 h per division, which is identical to the growth rate of the strain containing the pDW230 control plasmid. This suggests that no toxicity was observed for the overexpression of the GAL4DBD (Figure 2). The growth rates of the strains expressing GAL4DBD/SP1(1) (glutamine-rich), GAL4DBD/CTF (proline-rich) and GAL4DBD/VP16 (acidic-rich) were only 6.3, 6.8 and 8.3 h per division, respectively. In MMRT, no difference in growth rate was observed between the yeast containing the GAL4DBD expression plasmid and the yeast containing the GAL4DBD/VP16 expression plasmid. Therefore, the reduction in growth rate is due to the overexpression of the different GAL4DBD–TAD fusion proteins. These observations indicate that the tested GAL4DBD–TAD fusions are toxic when overexpressed in S.pombe. This toxicity may result from the squelching of components of the yeast transcriptional machinery by the overexpressed activation domains. Overexpression of acidic domains was also shown to be toxic in S.cerevisiae (Gill and Ptashne, 1988), and the screening for extragenic suppressors of this toxicity has allowed the isolation of transcriptional adaptors for acidic activation domains (Berger et al., 1992). Figure 2.Growth curve of S.pombe strain P2 expressing different GAL4DBD–TAD fusions. To induce the expression of the GAL4DBD–TAD fusions by the activation of the thiamine-repressible PHO4 promoter, cells were washed three times and put into culture in the absence of thiamine. In control samples (+ thia), the cells were grown in the presence of thiamine. At different times points, samples were taken and the cell number determined. The growth rate was estimated 35 h after induction. Download figure Download PowerPoint All three types of mammalian activation domains stimulate transcription in S.pombe in a position close to the TATA box The different GAL4DBD–TAD fusions were tested for transactivation in a proximal position in two reporter yeast strains, A1 and A2 (Figure 1). In both, one GAL4p 17mer recognition sequence (GAL4UAS) was inserted immediately upstream of the TATA box from the S.pombe ADH minimal (ADHmin) promoter. In reporter strain A1, this chimeric GAL4UAS–ADHmin promoter was cloned upstream of the neo gene, and the reporter construct was stably integrated into the ARG3 locus of the S.pombe genome. Transactivation in that reporter strain was determined by the relative G418 resistance level of the yeast transformants. Therefore, the yeast colonies were grown on minimal medium (MM) containing a limiting concentration of thiamine (5 μM) to repress the expression of the chimeric activators and replica-plated on YPD medium lacking thiamine and containing different concentrations of G418. In strain A2, the GAL4UAS–ADHmin chimeric promoter was cloned upstream of lacZ, and this fusion reporter gene was integrated into ADE6 locus. Transactivation of that reporter gene was determined by measuring β-galactosidase activity from yeast cells grown in liquid culture. β-Galactosidase activities were measured 8 h after shifting the culture from MM containing thiamine to MM lacking thiamine. Under these conditions, the different fusion proteins were produced in similar amounts, at least as determined by Western blot using GAL4 antibody (data not shown), and were not toxic for the cells (see Figure 2). The basal level of resistance for the A1 yeast is 100 mg of G418/l, which is equivalent to the resistance level observed for strain P2 that does not contain a neo reporter cassette (data not shown). The transformation of the GAL4DBD expression plasmid and the empty vector pDW230 as controls does not increase the resistance level of the A1 strain, indicating that no transactivation was observed (Figure 3A). In parallel, this was confirmed with the other test system by measurement of the β-galactosidase activity in the A2 strain (Figure 3B). Figure 3.Activation of transcription by the different GAL4DBD–TADs close to the TATA box and 87 bp upstream from the TATA box. Transcriptional activation of the reporter genes by the GAL4DBD–TADs placed close to the TATA box was estimated by the relative G418 resistance levels (A) in strain A1. Transcriptional activation by the GAL4DBD–TADs in a proximal context (solid bar) and 87 bp upstream of the TATA box (open bar) was determined by the relative β-galactosidase activities (B) in strains A2 and B, respectively. The values listed in (B) represent the β-galactosidase activities obtained with the different GAL4DBD–TADs after subtraction of the β-galactosidase background activity of strain A2 and B, respectively. Download figure Download PowerPoint The acidic-type GAL4DBD/VP16TAD activators yield high G418 resistance levels in strain A1 and high β-galactosidase levels in strain A2 (Figure 3A and B, respectively). These activations are comparable with the results obtained for the same fusion proteins expressed in S.cerevisiae and in mammalian cells (Seipel et al., 1992; Künzler et al., 1994). In addition, and like in mammalian cells (Seipel et al., 1992), we observed that the truncated version of VP16 TAD (amino acids 454–490) activates only half as potently as compared with the intact VP16 TAD (amino acids 413–490). The proline-rich activation domains of AP-2 and CTF also strongly stimulate transcription in S.pombe from close to the TATA box. In contrast to S.cerevisiae, the level of stimulation for the proline-rich and acidic-rich activation domains in S.pombe is equivalent, similar to what was found previously in mammalian cells. We also observed that the truncated version of AP-2 TAD (amino acids 31–76) is half as active as the intact AP-2 TAD (amino acids 31–117). This shorter AP-2 TAD was also less active in S.cerevisiae. Finally, the glutamine-rich activation domains of Sp1, Oct1 and Oct2 increase both the G-418 resistance levels of the A1 strain and the β-galactosidase activity of the A2 strain, indicating that these domains are functional in S.pombe. Therefore, unlike S.cerevisiae, the glutamine-rich activation domains stimulate transcription in S.pombe. We conclude from these results that S.pombe resembles, more so than S.cerevisiae, a mammalian cell with regard to the functionality of the different activation domains, at least when the latter operate close to the TATA-box. The three types of activation domains stimulate S.pombe transcription 87 bp upstream of the TATA box A 71 bp fragment of the luciferase gene was inserted between the GAL4UAS and the ADH minimal promoter, resulting in a final spacing of 87 bp. This chimeric promoter was cloned upstream of the lacZ gene and allowed the different TA domains to be tested in a more upstream context. As for the A2 strain, this reporter gene was also integrated at the ADE6 locus in the S.pombe genome, and this yeast reporter has been named strain B. β-Galactosidase activity was determined as for strain A2. The selected luciferase fragment does not contain any known binding site for yeast transcription factors, and the reporter strain B displays the same low β-galactosidase activity as the A2 strain (data not shown). The levels of transactivation observed for the acidic-rich TA domains in strain B are between 15 and 20% lower than the levels obtained in strain A2 (Figure 3B), indicating that a 87 bp upstream shift between the GAL4UAS and the TATA box had a limited impact on the level of transactivation by these domains in S.pombe. Similar observations were made for the proline-rich activation domains, with the exception of the CTF domain for which the level of activation drops by 40% in reporter B. Stimulation of transcription by the glutamine-rich domains seems to be more efficient in close proximity to the TATA box, because the insertion of an extra 87 bp between the TATA box and the GAL4UAS leads to a decrease in transcription activity of up to 50%. Therefore, when tested 87 bp usptream of the TATA box, all three types of activation domains still stimulate transcription in S.pombe. However, glutamine-rich TA domains seem to be more affected by the introduced 87 bp spacing than do the proline-rich and the acidic-rich TA domains. Only acidic-rich and proline-rich TA domains stimulate S.pombe transcription in a remote position 235 bp upstream of the TATA box To investigate the transactivation by these different TA domains in an enhancer-like position even more remote from the TATA box, we inserted 235 bp of spacer DNA derived from the luciferase open reading frame between the five copies of GAL4UAS and the ADHmin promoter. This 5GAL4UAS–ADHmin chimeric promoter was cloned upstream of the neo gene and, as in strain A1, this reporter was integrated at the ARG3 locus in the S.pombe genome, generating reporter strain C. Transactivation is documented by G418 resistance levels and Neo antigen levels (Figure 4A and B, respectively). Figure 4.Transcriptional activation by the different GAL4DBD–TADs remote (‘235 bp’ away) from the TATA box. Transcription activation by the GAL4DBD–TADs in strain C is documented in terms of G418 resistance levels (A) and Neo antigen levels (B). The values listed in (B) represent the Neo antigen levels obtained with the different GAL4DBD–TADs after subtraction of the Neo antigen background level of strain C. Download figure Download PowerPoint We observed a strong stimulation by the acidic TADs acting from the remote promoter position. As in the proximal context, we observed that the truncated TAD of VP16 is only half as potent as its intact counterpart. The proline-rich TADs can still stimulate transcription in S.pombe from a remote position. However, in contrast to the results which were observed at the proximal TATA box position, these domains are 12- to 25-fold less potent than the VP16 activation domain for activation of transcription from a ‘235 bp’ remote position. This differs from the results obtained in S.cerevisiae where the same proline-rich domains were inactive when tested in a similar ‘226 bp’ remote position (Künzler et al., 1994). Glutamine-rich activation domains fail to transactivate from a ‘235 bp’ remote position in S.pombe, except for the Oct2 domain, for which we observed a weak activation equivalent to the level obtained with the proline-rich activation domains. Therefore, as in mammalian cells, the glutamine-rich activation domains display very low transcriptional activity when tested in a context distal from the TATA box in S.pombe. In summary, all types of mammalian transactivation domain function in the fission yeast S.pombe similarly to how they do in mammalian cells but differently from how they do in the budding yeast S.cerevisiae. Discussion Striking similarity between mammalian cells and S.pombe with respect to stimulation of transcription by three classes of mammalian TAD In this study, we tested the three different types of TADs for their capacity to stimulate transcription in S.pombe from proximal and remote positions upstream of the TATA box. We have shown that acidic activation domains strongly stimulate transcription from both proximal or remote positions. These domains thus function similarly in S.pombe, in mammalian cells and in S.cerevisiae (Seipel et al., 1992; Künzler et al., 1994), indicating a conserved mechanism between mammalian cells and both yeast species. The proline-rich activation domains display strong activities when tested in a close proximal position to the TATA box in S.pombe. The levels of induction observed for these domains are comparable with the levels obtained with the acidic-rich domains, at least in the same promoter context. However, in contrast to the acidic-rich domains, the proline-rich domains stimulate transcription in a position remote (235 bp upstream) from the TATA box only weakly. Although these domains were shown to have considerable promoter activity and weak enhancer activity in mammalian cells (Seipel et al., 1992), they stimulate transcription only weakly from a position close to the TATA box in S.cerevisiae (Künzler et al., 1994). Finally, the glutamine-rich activation domains were shown to have strong promoter activities and no enhancer activity in mammalian cells (Seipel et al., 1992). In S.pombe, we also showed that these domains stimulate transcription only from a TATA box-proximal context, with the exception of the Oct2 TAD which is able to activate transcription weakly from a remote position. These glutamine-rich activation domains do not function in S.cerevisiae in any promoter context (Künzler et al., 1994), suggesting a divergence between the budding and the fission yeast for the utilization of glutamine-rich activation domains. Therefore, with respect to the use of the different types of activation domains, we conclude that S.pombe resembles mammalian cells more than S.cerevisiae does. Possible targets for the glutamine-rich activation domains in S.pombe All activation domains probably induce transcription by accelerating, during the assembly of the pre-initiation complex, the recruitment of GTFs such as TBP, TBP-associated factors (TAFs) and TFIIB (Zawel and Reinberg, 1995; Pugh, 1996), or by contacting the already partially pre-assembled RNA polymerase II holoenzyme (Koleske and Young, 1994). For example, it was shown previously that the glutamine-rich activation domains of Sp1 and Antennapedia (Antp) bind directly and specifically to the C-terminal domain of TBP, which is evolutionarily conserved from Drosophila to man (Emili et al., 1994). In addition, the ability of Sp1 activation domains to interact directly with the TBPs of various species correlates well with their ability to activate transcription in extracts derived from these species. In contrast, these activation domains interact weakly with the S.cerevisiae TBP (Emili et al., 1994). This weak interaction between TBP and the glutamine-rich domains may explain why these domains failed to stimulate transcription in S.cerevisiae. However, glutamine-rich activation domains still failed to activate tra" @default.
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• W2016070620 title "Three classes of mammalian transcription activation domain stimulate transcription in Schizosaccharomyces pombe" @default.
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