Structure-function analysis of the TBP-binding protein Dr1 reveals a mechanism for repression of class II gene transcription.

Yeung, Kam C.; Inostroza, J. A.; Mermelstein, Fred H.; Kannabiran, Chitra; Reinberg, Danny

doi:10.1101/gad.8.17.2097

Cited by 94 publications

(105 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…These results extend the known negative regulatory effects of NC2 on core RNA polymerase II in vitro (16,17,20,(42)(43)(44)(47)(48)(49)(50) and confirm that this regulator can inhibit transcription by RNA polymerase II in vivo.…”

Section: Discussionsupporting

confidence: 70%

See 1 more Smart Citation

Functional antagonism between RNA polymerase II holoenzyme and global negative regulator NC2 in vivo

Gadbois

Chao

Reese

et al. 1997

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

View full text Add to dashboard Cite

Activation of eukaryotic class II gene expression involves the formation of a transcription initiation complex that includes RNA polymerase II, general transcription factors, and SRB components of the holoenzyme. Negative regulators of transcription have been described, but it is not clear whether any are general repressors of class II genes in vivo. We reasoned that defects in truly global negative regulators should compensate for deficiencies in SRB4 because SRB4 plays a positive role in holoenzyme function. Genetic experiments reveal that this is indeed the case: a defect in the yeast homologue of the human negative regulator NC2 (Dr1⅐DRAP1) suppresses a mutation in SRB4. Global defects in mRNA synthesis caused by the defective yeast holoenzyme are alleviated by the NC2 suppressing mutation in vivo, indicating that yeast NC2 is a global negative regulator of class II transcription. These results imply that relief from repression at class II promoters is a general feature of gene activation in vivo.Activation of class II gene transcription in eukaryotes involves the recruitment of a transcription initiation complex that includes the RNA polymerase II holoenzyme (1-6). The yeast RNA polymerase II holoenzyme is a large multisubunit complex containing RNA polymerase II, a subset of the general transcription factors, and SRB regulatory proteins (7-11). Mammalian RNA polymerase II holoenzymes have also been purified, and an SRB7 homologue has been identified as a component of those complexes (12)(13)(14).For some class II genes, regulation appears to involve both positive and negative transcriptional regulators. The negative regulators that have been described include proteins purified for their ability to inhibit transcription in vitro (15-21) and genes identified because their products repress transcription from a subset of class II genes in vivo (21-29). For example, the human proteins NC1 (15, 16), NC2 or Dr1⅐DRAP1 (16,17,20), and DNA topoisomerase I (18, 19) repress basal transcription in vitro. The products of the yeast genes MOT1 (21-24), NOT1-4 (25-27), and SIN4 (28-29) negatively regulate at least a subset of yeast genes in vivo. Whether any of these negative regulators are generally employed for class II gene regulation in vivo is not yet clear.The RNA polymerase II C-terminal domain and the associated SRB complex have been implicated in the response to transcriptional activators (7-9, 30, 31). Two holoenzyme components, SRB4 and SRB6, have been shown to play essential and positive roles in transcription at the majority of class II genes in Saccharomyces cerevisiae (32). We reasoned that a defect in SRB4 might be alleviated by defects in general negative regulators and that knowledge of such regulators could contribute to our understanding of the mechanisms involved in gene regulation in vivo. Here we show that a deficiency in yeast NC2 can compensate for the global transcriptional defects caused by mutations in the SRB4 and SRB6 subunits of the RNA polymerase II holoenzyme and that NC2 is a globa...

show abstract

Section: Discussionsupporting

confidence: 70%

“…1b). A search of the GenBank database (June 7, 1996) revealed that the predicted protein has 39% identity over 99 aa to the NC2␣ (DRAP1) subunit of human NC2 (Dr1⅐DRAP1), which binds to TBP and represses transcription in vitro (16,17,20,(42)(43)(44)(47)(48)(49)(50). The gene encoding the putative yeast NC2␣ protein was named NCB1.…”

Section: Resultsmentioning

confidence: 99%

Functional antagonism between RNA polymerase II holoenzyme and global negative regulator NC2 in vivo

Gadbois

Chao

Reese

et al. 1997

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

View full text Add to dashboard Cite

show abstract

“…Even-skipped (Li and Manley, 1998), MOT1 (Auble and Hahn, 1993;Auble et al, 1994Auble et al, , 1997 and Dr1 (Han and Manley, 1993;Yeung et al, 1994) are examples of repressor proteins that interact with TBP, while KruÈ ppel and Engrailed (Zuo et al, 1991;Han and Manley, 1993;Licht et al, 1993) interact with TFIIE. RBP is a cellular repressor with speci®c DNA-binding activity which, in addition, contacts two coactivators (TAFII110 and TFIIA) and subverts activated transcription by interfering with the optimal interaction between these factors (Dou et al, 1994;Olave et al, 1998).…”

Section: Introductionmentioning

confidence: 99%

A murine ATFa-associated factor with transcriptional repressing activity

et al. 2000

View full text Add to dashboard Cite

The ATFa proteins, which are members of the CREB/ ATF family of transcription factors, have previously been shown to interact with the adenovirus E1a oncoprotein and to mediate its transcriptional activity; they heterodimerize with Jun, Fos or related transcription factors, possibly altering their DNA-binding speci®city; they also stably bind JNK2, a stress-induced protein kinase. Here we report the identi®cation and characterization of a novel protein isolated in a yeast two-hybrid screen using the N-terminal half of ATFa as a bait. This 1306-residue protein (mAM, for mouse ATFa-associated Modulator) is rather acidic (pHi 4.5) and contains high proportions of Ser/Thr (21%) and Pro (11%) residues. It colocalizes and interacts with ATFa in mammalian cells, contains a bipartite nuclear localization signal and possesses an ATPase activity. Transfection experiments show that mAM is able to downregulate transcriptional activity, in an ATPase-independent manner. Our results indicate that mAM interacts with several components of the basal transcription machinery (TFIIE and TFIIH), including RNAPII itself. Together, these ®ndings suggest that mAM may be involved in the ®ne-tuning of ATFaregulated gene expression, by interfering with the assembly or stability of speci®c preinitiation transcription complexes.

show abstract

“…The second region required for complete E1A repression activity, which includes residues 48 to 60, has a preponderance of acidic amino acids. This is unlike the repression subdomains of other direct repressors, such as Dr1, which contain few acidic amino acids and many alanine residues (57).…”

Section: Discussionmentioning

confidence: 99%

“…Several cellular cofactors have been reported recently to function as transcriptional repressors that interact with TBP, including Dr1, topoisomerase I, and MOT1 (for a review, see reference 60). Like E1A, Dr1 appears to block TFIIB interaction with TBP, and Dr1-mediated repression can be overcome by TBP (25,57). However, unlike E1A, Dr1 does not affect the binding of TBP to the TATA box.…”

Section: Discussionmentioning

confidence: 99%

The Adenovirus E1A Repression Domain Disrupts the Interaction between the TATA Binding Protein and the TATA Box in a Manner Reversible by TFIIB

Song

Loewenstein

Tóth

et al. 1997

Molecular and Cellular Biology

View full text Add to dashboard Cite

The human adenovirus E1A 243 amino acid oncoprotein possesses a transcription repression function that appears to be linked with its ability to induce cell cycle progression and to inhibit cell differentiation. The molecular mechanism of E1A repression has been poorly understood. Recently, we reported that the TATA binding protein (TBP) is a cellular target of E1A repression. Here we demonstrate that the interaction between TBP and the E1A repression domain is direct and specific. The TBP binding domain within E1A 243R maps to E1A N-terminal residues ϳ1 to 35 and is distinct from the TBP binding domain within conserved region 3 unique to the E1A 289R transactivator. An E1A protein fragment consisting of only the E1A N-terminal 80 amino acids (E1A 1-80) and containing the E1A repression function was found to block the interaction between TBP and the TATA box element as shown by gel mobility and DNase protection analysis. Interestingly, a preformed TBP-TATA box promoter complex can be dissociated by E1A 1-80. Further, TFIIB can prevent E1A disruption of TBP-TATA box interaction. TFIIB, like TBP, can overcome E1A repression of transcription in vitro. The ability of the E1A repression domain to block TBP interaction with the TATA box and the ability of TFIIB to reverse E1A disruption of the TBP-TATA box complex implies a mechanism for E1A repression distinct from those of known cellular repressors that target TBP.Adenovirus (Ad) encodes two major regulatory proteins of 243 and 289 amino acid residues (E1A 243R and E1A 289R for group C Ads) synthesized from alternatively spliced RNA transcripts of 12S and 13S encoded by early region E1A. E1A proteins contain multiple independent domains involved in diverse functions, including transcriptional activation, transcriptional repression, induction of cellular DNA synthesis, cell immortalization, and cell transformation as well as inhibition of metastasis and cell differentiation (for reviews, see references 9, 13, 43, and 47). E1A 289R differs from E1A 243R by conserved region 3 (CR3), a 46-amino-acid domain unique to 289R (see Fig. 1 for location of conserved regions). CR3 is essential (18,26,33,41,46) and sufficient (20, 34) for activation of viral early genes. Domains common to E1A 243R and 289R are required for the growth-regulatory functions of E1A; these include the relatively nonconserved N terminus (amino acid residues 1 to 39), CR1 (residues 40 to 80), and CR2 (residues 120 to 139) (40).Ad E1A 243R is a potent inducer of cellular DNA synthesis and a strong deregulator of cell cycle control. Two separate domains of E1A 243R can induce progression of quiescent cells to S-phase cellular DNA synthesis by independent pathways (24, 34). The first pathway involves binding sites within CR1 and CR2 for the retinoblastoma tumor suppressor protein, pRb. These sites sequester and dissociate pRb and related family members from complexes with the E2F family of transcription factors, whose activities can induce cell cycle progression (for a review, see reference 44). The second ...

show abstract

Structure-function analysis of the TBP-binding protein Dr1 reveals a mechanism for repression of class II gene transcription.

Cited by 94 publications

References 65 publications

Functional antagonism between RNA polymerase II holoenzyme and global negative regulator NC2 in vivo

Functional antagonism between RNA polymerase II holoenzyme and global negative regulator NC2 in vivo

A murine ATFa-associated factor with transcriptional repressing activity

The Adenovirus E1A Repression Domain Disrupts the Interaction between the TATA Binding Protein and the TATA Box in a Manner Reversible by TFIIB

Contact Info

Product

Resources

About