Solution Structure of the KIX Domain of CBP Bound to the Transactivation Domain of CREB: A Model for Activator:Coactivator Interactions

Radhakrishnan, Ishwar; Pérez-Alvarado, Gabriela C.; Parker, David B.; Dyson, H. Jane; Montminy, Marc; Wright, Peter E.

doi:10.1016/s0092-8674(00)80463-8

Cited by 703 publications

(998 citation statements)

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“…6). More generally, the fold of Med25 ACID is unique amongst known activator target domains 21,[45][46][47] such as Tfb1, Mdm2, PAS-B and the CBP KIX domain, which are all unrelated in structure (Fig. 6).…”

Section: Discussionmentioning

confidence: 99%

Structure and VP16 binding of the Mediator Med25 activator interaction domain

Vojnic

Mourão

Seizl

et al. 2011

Nat Struct Mol Biol

111

161

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4 0 4 VOLUME 18 NUMBER 4 APRIL 2011 nAture structurAl & moleculAr biologyActivator proteins regulate eukaryotic RNA polymerase II (Pol II) transcription in response to developmental and environmental signals. They bind to the DNA recognition sites of target genes with a sequence-specific DNA-binding domain, and recruit components of the Pol II machinery through a transcriptional activation domain (TAD) 1,2 . Activators have been classified as acidic, glutamine-rich, proline-rich and serine/threonine-rich, depending on the preponderance of amino acids in their TAD 3 . An archetypal acidic activator is the herpes simplex virus protein 16 (VP16), which activates the expression of immediate early viral genes during infection 4-8 . VP16 exerts its activating function through a C-terminal TAD that includes residues 413-490 (refs. 9-13). The VP16 TAD has been intensely used for studying transcriptional activation. Usually, the VP16 TAD is fused with its N terminus to the DNA-binding domain of the yeast transcription factor Gal4. The resulting Gal4-VP16 activator fusion protein stimulates transcription from promoters that contain Gal4-binding sites in the yeast and mammalian transcription systems in vivo 14,15 and in vitro 16,17 . This indicates that basic mechanisms of transcription activation are conserved amongst eukaryotes. The VP16 TAD targets basal Pol II transcription factors, including TFIIA, TFIIB, TFIID, the TFIIH subunit Tfb1/p62 (yeast/human) and the Mediator co-activator [18][19][20][21][22][23] . The VP16 TAD binds the Mediator subunit Med25 (also called Arc92) [24][25][26][27] , which is specific to higher eukaryotes. Med25 consists of two domains, the activator interaction domain (ACID) 24 that binds the VP16 TAD, and a 'von Willebrand factor type A' domain that anchors Med25 to Mediator 24 . Mediator generally conveys regulatory information by forming a bridge between activators and the basal Pol II machinery 28,29 . Whereas information on the core Mediator structure is emerging, structural information about its more peripheral activator-binding domains is limited to the KIX domain in subunit Med15 (or Arc105) 30,31 .The VP16 TAD is intrinsically flexible, whereas the VP16 core domain (residues 49-402) forms a stable structure 23,[32][33][34] . The TAD contains two functional subdomains, H1 (residues 410-452) and H2 (residues 453-490), which activate transcription independently 35,36 . Both H1 and H2 contain acidic amino acids, but specific bulky hydrophobic and aromatic residues are required for their function [36][37][38][39] . H2 has been proposed to adopt an α-helical conformation when bound to TFIIB 40 . NMR analysis revealed a nine-residue amphipathic α-helix in H2 that docks onto a PH fold in Tfb1 21 . On the basis of these and other results it is assumed that acidic TADs are flexible in their free state, but become transiently structured upon interaction with their target proteins.Here we report the NMR structure of the Mediator Med25 ACID and analyze its structural and functional interaction wi...

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Section: Discussionmentioning

confidence: 99%

Structure and VP16 binding of the Mediator Med25 activator interaction domain

Vojnic

Mourão

Seizl

et al. 2011

Nat Struct Mol Biol

111

161

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“…One protein that follows the folding after binding pathway is the pKID of the transcription factor CREB. pKID exists as an unfolded protein in solution in vitro (Radhakrishnan et al 1997) but folds upon binding to the KIX domain of the CREB binding protein (Parker et al 1996). The structure of the pKID-KIX complex has been solved by NMR (Radhakrishnan et al 1997), revealing that the pKID forms two helices and that the C-terminal helix in particular is very well-defined, interacting specifically with KIX (Fig.…”

Section: Coupled Folding/bindingmentioning

confidence: 99%

Protein dynamics and function from solution state NMR spectroscopy

Kovermann

Rogne

Wolf‐Watz

2016

Quart. Rev. Biophys.

143

122

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Abstract. It is well-established that dynamics are central to protein function; their importance is implicitly acknowledged in the principles of the Monod, Wyman and Changeux model of binding cooperativity, which was originally proposed in 1965. Nowadays the concept of protein dynamics is formulated in terms of the energy landscape theory, which can be used to understand protein folding and conformational changes in proteins. Because protein dynamics are so important, a key to understanding protein function at the molecular level is to design experiments that allow their quantitative analysis. Nuclear magnetic resonance (NMR) spectroscopy is uniquely suited for this purpose because major advances in theory, hardware, and experimental methods have made it possible to characterize protein dynamics at an unprecedented level of detail. Unique features of NMR include the ability to quantify dynamics (i) under equilibrium conditions without external perturbations, (ii) using many probes simultaneously, and (iii) over large time intervals. Here we review NMR techniques for quantifying protein dynamics on fast (ps-ns), slow (μs-ms), and very slow (s-min) time scales. These techniques are discussed with reference to some major discoveries in protein science that have been made possible by NMR spectroscopy.

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“…Since the interaction between MBF1 and FTZ-F1 is rather stable, it is implied that the N-terminal residues undergo folding transitions when they encounters the activator. Induced folding from otherwise unfolded states is often encountered in protein-protein interactions between transcriptional factors: the activation domain of p53 by binding to MDM2 (Kussie et al 1996), the activation domain of VP16 by binding to hTAF II 31 (Uesugi et al 1997), KIX domain by binding to CBP (Radhakrishnan et al 1997), and the transcriptional inhibitory domain of dTAF II 230 by binding to TBP (Liu et al 1998). MBF1 is characterized by its ability to bind several different transcriptional factors, for instance: human MBF1 can bind to either Ad4BP, ATF1, CREB1 or CREBP1 (Y. Kabe, M. Goto, D. Shima, T. Wada, K. Morohashi, M. Shirakawa, S. Hirose & H. Handa, unpublished result).…”

Section: The N-terminal Residuesmentioning

confidence: 99%