Structural analyses of CREB-CBP transcriptional activator-coactivator complexes by NMR spectroscopy: implications for mapping the boundaries of structural domains

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

doi:10.1006/jmbi.1999.2658

Cited by 69 publications

(76 citation statements)

References 23 publications

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“…2B, lane 9). It has been reported that the structure of the KIX domain is disordered when the ␣-3 helix is deleted (39), so it is possible that the MLL activation domain recognizes an aspect of the structure formed by the three helices and not necessarily the third helix itself. Therefore, we conclude that the minimal regions mediating the MLL-CBP interaction correspond to CBP amino acids 581 to 687 and MLL amino acids 2829 to 2883.…”

Section: Resultsmentioning

confidence: 99%

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MLL and CREB Bind Cooperatively to the Nuclear Coactivator CREB-Binding Protein

Ernst

Wang

Huang

et al. 2001

Molecular and Cellular Biology

224

227

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A fragment of the mixed-lineage leukemia (MLL) gene (Mll, HRX, ALL-1) was identified in a yeast genetic screen designed to isolate proteins that interact with the CREB-CREB-binding protein (CBP) complex. When tested for binding to CREB or CBP individually, this MLL fragment interacted directly with CBP, but not with CREB. In vitro binding experiments refined the minimal region of interaction to amino acids 2829 to 2883 of MLL, a potent transcriptional activation domain, and amino acids 581 to 687 of CBP (the CREB-binding or KIX domain). The transactivation activity of MLL was dependent on CBP, as either adenovirus E1A expression, which inhibits CBP activity, or alteration of MLL residues important for CBP interaction proved effective at inhibiting MLL-mediated transactivation. Single amino acid substitutions within the MLL activation domain revealed that five hydrophobic residues, potentially forming a hydrophobic face of an amphipathic helix, were critical for the interaction of MLL with CBP. Using purified components, we found that the MLL activation domain facilitated the binding of CBP to phosphorylated CREB. In contrast with paradigms in which factors compete for limiting quantities of CBP, these results reveal that two distinct transcription factor activation domains can cooperatively target the same motif on CBP.The interaction of the cyclic AMP response element-binding protein (CREB) with its coactivator, CREB-binding protein (CBP), is one of the best characterized signal-regulated activation domain-coactivator interactions. CREB is a signal-dependent transactivator of the bZIP family which becomes phosphorylated on serine 133 in response to stimuli that result in an increase in intracellular cyclic AMP or Ca 2ϩ (33). CBP, as well as its homologue p300, binds to CREB upon phosphorylation at serine 133 within the kinase-inducible activation domain of CREB (8,32). CBP recruitment by phosphorylated CREB results in rapid induction of gene expression that is mediated both by direct recruitment of the basal transcription machinery and the intrinsic acetyltransferase activity of CBP (26,33). A vast number of signal-responsive and developmentally regulated transcription factors interact with different regions of CBP and p300. The CREB-binding (or KIX) domain alone interacts with at least 10 distinct families of transcriptional activators (17). Despite the number of well-established protein interactions involving CBP and p300, the only complex for which structural information is available is the phosphorylated CREB-CBP KIX domain interaction (38, 39).The solution structure of the phosphorylated CREB-CBP KIX complex revealed that the activation domain of CREB undergoes a dramatic random coil-to-helix transition upon binding to the KIX domain (38). Conversely, the ␣-3 helix of the KIX domain undergoes a more subtle change upon binding to phosphorylated CREB. Two lines of evidence suggest that this binding event may be regulated at a level beyond serine 133 phosphorylation. First, CREB phosphorylation has been noted...

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Section: Resultsmentioning

confidence: 99%

“…The CREB-binding (or KIX) domain alone interacts with at least 10 distinct families of transcriptional activators (17). Despite the number of well-established protein interactions involving CBP and p300, the only complex for which structural information is available is the phosphorylated CREB-CBP KIX domain interaction (38,39).…”

mentioning

confidence: 99%

MLL and CREB Bind Cooperatively to the Nuclear Coactivator CREB-Binding Protein

Ernst

Wang

Huang

et al. 2001

Molecular and Cellular Biology

224

227

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“…Contained within CBP is an 87-residue-long domain called kinase-inducible domain interacting (KIX) that is critical for regulating transcriptional activity (for review, see ref. 11) and can be cooperatively targeted by a diverse set of transcription factors (12)(13)(14)(15)(16)(17) and small molecules (18)(19)(20)(21)(22)(23). The structure of KIX ( Fig.…”

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confidence: 99%

Prepaying the entropic cost for allosteric regulation in KIX

Law¹,

Gagnon²,

Mapp

et al. 2014

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

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The kinase-inducible domain interacting (KIX) domain of the CREB binding protein (CBP) is capable of simultaneously binding two intrinsically disordered transcription factors, such as the mixedlineage leukemia (MLL) and c-Myb peptides, at isolated interaction sites. In vitro, the affinity for binding c-Myb is approximately doubled when KIX is in complex with MLL, which suggests a positive cooperative binding mechanism, and the affinity for MLL is also slightly increased when KIX is first bound by c-Myb. Expanding the scope of recent NMR and computational studies, we explore the allosteric mechanism at a detailed molecular level that directly connects the microscopic structural dynamics to the macroscopic shift in binding affinities. To this end, we have performed molecular dynamics simulations of free KIX, KIX-c-Myb, MLL-KIX, and MLL-KIX-c-Myb using a topology-based Go-like model. Our results capture an increase in affinity for the peptide in the allosteric site when KIX is prebound by a complementary effector and both peptides follow an effector-independent folding-and-binding mechanism. More importantly, we discover that MLL binding lowers the entropic cost for c-Myb binding, and vice versa, by stabilizing the L 12 -G 2 loop and the C-terminal region of the α 3 helix on KIX. This work demonstrates the importance of entropy in allosteric signaling between promiscuous molecular recognition sites and can inform the rational design of small molecule stabilizers to target important regions of conformationally dynamic proteins.

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“…Many models of long-term memory storage suggest that the phosphorylation of a constitutively present factor such as CREB may mediate the induction of gene expression required for memory consolidation. The importance of understanding transcription factor-coactivator interactions is underscored by the numerous studies examining the KID-KIX interaction (Parker et al 1998Radhakrishnan et al 1998Radhakrishnan et al , 1999Shaywitz et al 2000;Campbell and Lumb 2002), including a solution structure of the transactivation domain of CREB in a complex with the CBP KIX domain (Radhakrishnan et al 1997). In contrast to our understanding of the biochemical details regarding the KID-KIX interaction, the function of the CBP KIX domain in vivo is not known.…”

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confidence: 99%

A transcription factor-binding domain of the coactivator CBP is essential for long-term memory and the expression of specific target genes

Wood

Attner²,

Oliveira³

et al. 2006

Learn. Mem.

176

163

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Transcriptional activation is a key process required for long-term memory formation. Recently, the transcriptional coactivator CREB-binding protein (CBP) was shown to be critical for hippocampus-dependent long-term memory and hippocampal synaptic plasticity. As a coactivator with intrinsic histone acetyltransferase activity, CBP interacts with numerous transcription factors and contains multiple functional domains. Currently, it is not known which transcription factor-binding domain of CBP is essential for memory storage. Using mice that carry inactivating mutations in the CREB-binding (KIX) domain of the coactivator CBP (CBP KIX/KIX mice), we show that the KIX domain is required for long-term memory storage. These results are the first to identify an in vivo function for the KIX domain of CBP in the brain, and they suggest that KIX-interacting transcription factors recruit CBP histone acetyltransferase activity during long-term memory storage. One such KIX-interacting factor is the transcription factor CREB. Using quantitative real-time RT-PCR, we find that the expression of specific CREB target genes is reduced in the hippocampi of CBP KIX/KIX mice during memory consolidation. The recruitment of the transcriptional coactivator CBP via the KIX domain thus imparts target gene-dependent selectivity to CREB-driven transcriptional regulation, thereby activating genes required for the long-term storage of hippocampus-dependent memory.Recently, we and others have shown that CREB-binding protein (CBP) is critical for hippocampus-dependent long-term memory (Oike et al. 1999;Bourtchouladze et al. 2003;Korzus et al. 2004) and synaptic plasticity (Alarcon et al. 2004;Wood et al. 2005; for review, see Josselyn 2005). CBP is a transcriptional coactivator that regulates transcription via direct interactions with transcription factors and the basal transcription machinery as well as via its intrinsic histone acetyltransferase (HAT) activity, which modifies chromatin structure (Goodman and Smolik 2000;Kalkhoven 2004). CBP interacts with numerous transcription factors and contains multiple functional domains. Studies focusing on memory storage have demonstrated that the CBP HAT domain is necessary for memory storage (Korzus et al. 2004) and suggested that CBP is recruited to DNA by transcription factors interacting with the amino terminus of CBP (Oike et al. 1999;Bourtchouladze et al. 2003;Wood et al. 2005). However, which transcription factor-binding domain of CBP is essential for memory storage remains a key open question.The KIX domain of CBP provides a critical interface for the recruitment of CBP by specific transcription factors and the subsequent regulation of gene expression. Several transcriptional activators bind to the KIX domain of CBP, but most of these factors do so in a constitutive fashion. The interactions between CREB and CBP, however, are regulated by the phosphorylation state of CREB. Phosphorylation of CREB at Ser 133 in the kinaseinducible domain (KID), as a result of multiple signaling transduction casca...

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Structural analyses of CREB-CBP transcriptional activator-coactivator complexes by NMR spectroscopy: implications for mapping the boundaries of structural domains

Cited by 69 publications

References 23 publications

MLL and CREB Bind Cooperatively to the Nuclear Coactivator CREB-Binding Protein

MLL and CREB Bind Cooperatively to the Nuclear Coactivator CREB-Binding Protein

Prepaying the entropic cost for allosteric regulation in KIX

A transcription factor-binding domain of the coactivator CBP is essential for long-term memory and the expression of specific target genes

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