Region of Yeast TAF 130 Required for TFIID To Associate with Promoters

Mencı́a, Mario; Struhl, Kevin

doi:10.1128/mcb.21.4.1145-1154.2001

Cited by 18 publications

(24 citation statements)

References 63 publications

(112 reference statements)

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“…Portions of the central conserved domain are deleted in mutants ⌬5-to ⌬13Taf1p, thus suggesting that this HAT function could be one of the nonscaffold functions of Taf1p. The Taf1p domain defined by ⌬16 has been implicated in promoter (DNA) binding by others (44). Taken together, these results underscore the modular nature of this protein.…”

Section: Discussionsupporting

confidence: 58%

“…Comparable detailed protein-protein interaction studies with metazoan Taf1p have not yet been reported; thus, it remains to be determined whether a similar (58,60). In this regard, it is notable that Cterminal truncation mutants of Taf1p that contain region 4 sequences have been found to form TFIID-like subcomplexes (18,44), though in these experiments not all the TFIID-Tafps were probed for. In light of our data, we assume that region 4 most likely contributed critically to the assembly of these TFIID subcomplexes.…”

Section: Discussionmentioning

confidence: 97%

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Molecular and Genetic Characterization of a Taf1p Domain Essential for Yeast TFIID Assembly

Singh

Bland

Weil

2004

Molecular and Cellular Biology

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Yeast Taf1p is an integral component of the multiprotein transcription factor TFIID. By using coimmunoprecipitation assays, coupled with a comprehensive set of deletion mutants encompassing the entire open reading frame of TAF1, we have discovered an essential role of a small portion of yeast Taf1p. This domain of Taf1p, termed region 4, consisting of amino acids 200 to 303, contributes critically to the assembly and stability of the 15-subunit TFIID holocomplex. Region 4 of Taf1p is mutationally sensitive, can assemble several Tafps into a partial TFIID complex, and interacts directly with Taf4p and Taf6p. Mutations in Taf1p-region 4 induce temperature-conditional growth of yeast cells. At the nonpermissive temperature these mutations have drastic effects on both TFIID integrity and mRNA synthesis. These data are consistent with the hypothesis that Taf1p subserves a critical scaffold function within the TFIID complex. The significance of these data with regard to TFIID structure and function is discussed.TFIID, one of several multiprotein general transcription factors required for RNA polymerase II (PolII)-catalyzed mRNA gene transcription, is comprised of the TATA-binding protein (TBP) and 14 distinct TBP-associated factors (TAFs or Tafps) that range in M r from 17,000 to 150,000 in Saccharomyces cerevisiae (62,63,65,75). These TFIID subunits share significant sequence conservation with their metazoan orthologs (1,22,26). In vitro, PolII needs six general transcription factors, TFIIA, -B, -D, -E, -F, and -H, to form functional preinitiation complexes (PICs) (12, 51, 54); TFIID plays several critical roles in this process. First, TFIID functions during promoter recognition by binding directly to the promoter (67,76,77). On promoters containing a TATA box, Tafps help stabilize TBP binding to the TATA element via interaction with TATA-proximal initiator DNA sequences (69). Tafp-DNA interactions may be particularly important for TFIID binding to promoters that lack a canonical TATA element and contain TATA-proximal initiator and distal promoter element sequences (13,15,42,69,76). Second, Tafps can function as coactivators during transcriptional activation by making direct contacts with specific activators leading to an increase in PolII PIC formation (1, 16). Finally, Taf1p contains several distinct enzymatic activities, those of histone acetyltransferase (HAT) (46), protein kinase (19), and ubiquitin ligase (55). These enzymatic activities presumably modify proteins that stimulate PIC formation and/or function, leading to PolII transcription initiation.In most contexts, the Tafp subunits of TFIID are essential for survival, as Tafp inactivation or depletion results in cessation of specific mRNA synthesis and loss of cell viability (3,4,33,34,56,59,64,65,78). Because of these critical roles, the composition, organization, assembly, structure, and function of the TFIID complex have been topics of great interest. TBP appears to be incorporated into the TFIID complex primarily through its interaction with the biparti...

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Section: Discussionsupporting

confidence: 58%

Section: Discussionmentioning

confidence: 97%

Molecular and Genetic Characterization of a Taf1p Domain Essential for Yeast TFIID Assembly

Singh

Bland

Weil

2004

Molecular and Cellular Biology

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“…Transcription from this altered promoter depended heavily on TAF1 and TAF4, but not on TAF5, TAF6, or TAF9. Previous studies identified a conserved C-terminal region of yeast TAF1 that was required for association of TFIID with endogenous promoters (23). Perhaps this same region of Drosophila TAF1 is required for TFIID to bind efficiently to TATA-less, DPE-containing promoters but is dispensable at promoters containing a TATA box.…”

Section: Discussionmentioning

confidence: 99%

TAF4 nucleates a core subcomplex of TFIID and mediates activated transcription from a TATA-less promoter

Wright

Marr

Tjian

2006

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

137

146

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Activator-dependent recruitment of TFIID initiates formation of the transcriptional preinitiation complex. TFIID binds core promoter DNA elements and directs the assembly of other general transcription factors, leading to binding of RNA polymerase II and activation of RNA synthesis. How TATA box-binding protein (TBP) and the TBP-associated factors (TAFs) are assembled into a functional TFIID complex with promoter recognition and coactivator activities in vivo remains unknown. Here, we use RNAi to knock down specific TFIID subunits in Drosophila tissue culture cells to determine which subunits are most critical for maintaining stability of TFIID in vivo. Contrary to expectations, we find that TAF4 rather than TBP or TAF1 plays the most critical role in maintaining stability of the complex. Our analysis also indicates that TAF5, TAF6, TAF9, and TAF12 play key roles in stability of the complex, whereas TBP, TAF1, TAF2, and TAF11 contribute very little to complex stability. Based on our results, we propose that holo-TFIID comprises a stable core subcomplex containing TAF4, TAF5, TAF6, TAF9, and TAF12 decorated with peripheral subunits TAF1, TAF2, TAF11, and TBP. Our initial functional studies indicate a specific and significant role for TAF1 and TAF4 in mediating transcription from a TATA-less, downstream core promoter element (DPE)-containing promoter, whereas a TATA-containing, DPE-less promoter was far less dependent on these subunits. In contrast to both TAF1 and TAF4, RNAi knockdown of TAF5 had little effect on transcription from either class of promoter. These studies significantly alter previous models for the assembly, structure, and function of TFIID.RNA interference ͉ TATA box-binding protein ͉ S2 cells R egulated initiation of transcription to produce mRNA in eukaryotes requires the stepwise assembly of an elaborate multiprotein preinitiation complex consisting of the general transcription factors, various coactivators, and RNA polymerase II (for review, see ref. 1). The core promoter-recognition complex, TFIID, consists of the TATA box-binding protein (TBP) and 8-12 TBP-associated factors (TAFs). In addition to binding core promoter elements and initiating formation of the preinitiation complex, this TBP⅐TAF multisubunit transcription factor also serves as a coactivator by transmitting signals from sequence-specific activators to other components of the basal machinery (for review, see ref.2).Critical to dissecting the diverse functions of TFIID in both promoter recognition and coactivation is an understanding of how the complex is assembled and maintained in cells. Initial in vitro assembly reactions suggested that TBP and the largest TAF subunit (TAF1) may form a scaffold on which the other TAFs bind to form holo-TFIID (3). Subsequent studies have proposed that TAF5 may dimerize and also help coordinate complex assembly (4). Recent low-resolution electron microscopy͞single-particle reconstruction models of TFIID have revealed a trilobed architecture containing a large central cavity that has been conse...

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“…16 and 17). Like its metazoan counterparts, the yeast protein, TAF130p, contains a histone acetyltransferase domain as well as a number of other essential sequences (18,19) whose exact functions remain to be defined. One highly conserved and important element found in this (family of) TAF are domains capable of directly binding the TBP subunit of TFIID (17, 18, 20 -24).…”

mentioning

confidence: 99%

Fluorescence-based Analyses of the Effects of Full-length Recombinant TAF130p on the Interaction of TATA Box-binding Protein with TATA Box DNA

Banik

Beechem²,

Klebanow³

et al. 2001

Journal of Biological Chemistry

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We have used a combination of fluorescence anisotropy spectroscopy and fluorescence-based native gel electrophoresis methods to examine the effects of the transcription factor IID-specific subunit TAF130p (TAF145p) upon the TATA box DNA binding properties of TATA box-binding protein (TBP). Purified full-length recombinant TAF130p decreases TBP-TATA DNA complex formation at equilibrium by competing directly with DNA for binding to TBP. Interestingly, we have found that full-length TAF130p is capable of binding multiple molecules of TBP with nanomolar binding affinity. The biological implications of these findings are discussed.Eukaryotic DNA-dependent RNA polymerase II works in concert with the six general transcription factors (GTFs) 1 TFIIA, -B, -D, -E, -F, and -H to catalyze mRNA gene transcription (1). These components act either sequentially (2) or as a part of a holoenzyme complex (3, 4) to form a multicomponent preinitiation complex on promoter DNA. A highly conserved feature of most eukaryotic mRNA promoters is the TATA box element present 25 base pairs upstream from the transcription start site. TFIID, in combination with TFIIA and TFIIB, can recognize the TATA element and form a platform for subsequent preinitiation complex formation. The TATA box-binding protein (TBP), as its name suggests, is the protein within the 15-subunit TFIID holocomplex (5) that makes primary contact with the TATA element, though several TBP-associated factor (TAF) subunits comprising TFIID contribute to promoter binding (6 -8). Binding of TFIID to TATA elements is central to the control of transcription (9, 10).Recruiting TFIID to the promoter, a process thought to be mediated in part by direct activator-TFIID interactions, is probably a key and universal mechanism of gene activation (9, 10). However, the largest subunit of metazoan TFIID, which in Drosophila (d) or humans (h) exhibits an apparent molecular mass of ϳ250 kDa, termed d-or hTAF II 250, respectively (herein termed d/hTAF250p), also contains a number of intrinsic enzymatic activities including histone acetyltransferase (11), protein kinase (12), and ubiquitin activating/conjugating activity (13). Each of these activities could be targets for transactivators, and mutation of histone acetyltransferase, protein kinase, or ubiquitin activating/conjugating activity domains decreases transcription of subsets of genes in vivo (13-15).The yeast ortholog of d/hTAF250, TAF130p, is encoded by a single-copy essential gene (TAF130/TAF145; Refs. 16 and 17). Like its metazoan counterparts, the yeast protein, TAF130p, contains a histone acetyltransferase domain as well as a number of other essential sequences (18, 19) whose exact functions remain to be defined. One highly conserved and important element found in this (family of) TAF are domains capable of directly binding the TBP subunit of TFIID (17, 18, 20 -24). Both Drosophila TAF250p and yeast TAF130p appear to contain at least two TBP binding domains (18,23,25): a high affinity N-terminal TBP binding domain and a less well de...

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Region of Yeast TAF 130 Required for TFIID To Associate with Promoters

Cited by 18 publications

References 63 publications

Molecular and Genetic Characterization of a Taf1p Domain Essential for Yeast TFIID Assembly

Molecular and Genetic Characterization of a Taf1p Domain Essential for Yeast TFIID Assembly

TAF4 nucleates a core subcomplex of TFIID and mediates activated transcription from a TATA-less promoter

Fluorescence-based Analyses of the Effects of Full-length Recombinant TAF130p on the Interaction of TATA Box-binding Protein with TATA Box DNA

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