Variable Control of Ets-1 DNA Binding by Multiple Phosphates in an Unstructured Region

Pufall, Miles A.; Lee, Gregory M.; Nelson, Mary L.; Kang, Hyun-Seo; Vėlyvis, Algirdas; Kay, Lewis E.; McIntosh, Lawrence P.; Graves, Barbara J.

doi:10.1126/science.1111915

Cited by 235 publications

(316 citation statements)

References 30 publications

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“…Multiple phosphorylated sites of the intrinsically disordered cyclin-dependent kinase inhibitor Sic1 bind a single site on the Cdc4 component of SCF ubiquitin ligase, with the dynamic equilibrium of this interaction allowing highaffinity switch-like binding 43 . In contrast, the intrinsically disordered N-terminal segment of the Ets-1 transcription factor has multiple phosphorylation sites that act as a rheostat to control autoinhibition of the ETS domain 44 . The R region may be a similar rheostat, except that primarily its nonphosphorylated state binds.…”

Section: Two Predominant Mechanismsmentioning

confidence: 99%

CFTR regulatory region interacts with NBD1 predominantly via multiple transient helices

Baker

Hudson

Kanelis

et al. 2007

Nat Struct Mol Biol

267

327

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The regulatory (R) region of the cystic fibrosis transmembrane conductance regulator (CFTR) is intrinsically disordered and must be phosphorylated at multiple sites for full CFTR channel activity, with no one specific phosphorylation site required. In addition, nucleotide binding and hydrolysis at the nucleotide-binding domains (NBDs) of CFTR are required for channel gating. We report NMR studies in the absence and presence of NBD1 that provide structural details for the isolated R region and its interaction with NBD1 at residue-level resolution. Several sites in the R region with measured fractional helical propensity mediate interactions with NBD1. Phosphorylation reduces the helicity of many R-region sites and reduces their NBD1 interactions. This evidence for a dynamic complex with NBD1 that transiently engages different sites of the R region suggests a structural explanation for the dependence of CFTR activity on multiple PKA phosphorylation sites.The CFTR chloride channel, the protein mutated in cystic fibrosis, is a member of the ATPbinding cassette (ABC) superfamily of proteins 1 . Like other members of the superfamily, CFTR has two membrane-spanning domains (MSD1 and MSD2) and two nucleotidebinding domains (NBD1 and NBD2). Intracellular regions between the transmembrane segments probably adopt helical structures that extend from the MSDs 2 . Unique to CFTR is the cytoplasmic intrinsically disordered 3,4 R region, of approximately 200 residues, which we refer to as a region rather than as a domain to reflect its lack of a stable, folded globular structure.

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Section: Two Predominant Mechanismsmentioning

confidence: 99%

CFTR regulatory region interacts with NBD1 predominantly via multiple transient helices

Baker

Hudson

Kanelis

et al. 2007

Nat Struct Mol Biol

267

327

View full text Add to dashboard Cite

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“…Multi-site phosphorylation has been documented for several factors, including Sic1 (Nash et al, 2001), NFAT (Okamura et al, 2000), Stat6 (Maiti et al, 2005) and Ets-1 (Pufall et al, 2005). It has been suggested to overcome a threshold needed to establish a biological response (Nash et al, 2001;Gunawardena, 2005;Pufall et al, 2005).…”

Section: Discussionmentioning

confidence: 99%

“…It has been suggested to overcome a threshold needed to establish a biological response (Nash et al, 2001;Gunawardena, 2005;Pufall et al, 2005). That this also may apply to Oct-1 is reflected by the additive effects of the sites within the A, B and C subregions.…”

Section: Discussionmentioning

confidence: 99%

DNA-PK phosphorylation sites on Oct-1 promote cell survival following DNA damage

et al. 2007

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“…This finding is in agreement with the major role of Ets-1 as an activator of Stromelysin-1 expression through its binding to the palindromic head-to-head EBS present in the promoter. Indeed, CaMKII phosphorylation of Ets-1 on residues Ser-251, Ser-270, Ser-273, Ser-282 and Ser-285 was reported to negatively regulate Ets-1 DNA binding in vitro and in vivo by reinforcing autoinhibition (Pufall et al, 2005). We previously demonstrated that reinforcing autoinhibition through CaMKII phosphorylation reduced dramatically the cooperative binding of Ets-1 to the EBS palindrome preventing the formation of the ternary Ets-1-DNA-Ets-1 complex, important for a full transactivation of the Stromelysin-1 promoter (Baillat et al, 2002).…”

Section: Stromelysin-1 Promoter Regulation By Endogenous Ets-1 D Bailmentioning

confidence: 99%

Stromelysin-1 expression is activated in vivo by Ets-1 through palindromic head-to-head Ets binding sites present in the promoter

et al. 2006

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Regulation of the gene expression of Stromelysin-1 (matrix metalloproteinase-3), a member of the matrix metalloproteinase family, is critical for tissue homeostasis. The Stromelysin-1 promoter is known to be transactivated by Ets proteins through palindromic headto-head Ets binding sites (EBS), an unusual configuration among metalloproteinase promoters. Patterns of increased co-expression of Stromelysin-1 and Ets-1 genes have been observed in pathological processes such as rheumatoid arthritis, glomerulonephritis and tumor invasion. In this context, we show in a synovial fibroblastic model cell line (HIG-82), which is able to co-express Stromelysin-1 and Ets-1, that the EBS palindrome is essential for the expression of Stromelysin-1. More precisely, using electrophoretic mobility shift assays, DNA affinity purification and chromatin immunoprecipitation, we demonstrate that endogenous Ets-1, but not Ets-2, is present on this palindrome. The use of a dominant-negative form of Ets-1 and the decrease of Ets-1 amount either by fumagillin, an antiangiogenic compound, or by short interfering RNA show that the activation rate of the promoter and the expression of Stromelysin-1 correlate with the level of endogenous Ets-1. Thus, it is the first demonstration, using this cellular model, that endogenously expressed Ets-1 is actually a main activator of the Stromelysin-1 promoter through its effective binding to the EBS palindrome.

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Variable Control of Ets-1 DNA Binding by Multiple Phosphates in an Unstructured Region

Cited by 235 publications

References 30 publications

CFTR regulatory region interacts with NBD1 predominantly via multiple transient helices

CFTR regulatory region interacts with NBD1 predominantly via multiple transient helices

DNA-PK phosphorylation sites on Oct-1 promote cell survival following DNA damage

Stromelysin-1 expression is activated in vivo by Ets-1 through palindromic head-to-head Ets binding sites present in the promoter

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