Interleukin-1α (IL-1α) is a proinflammatory cytokine and a key player in host immune responses in higher eukaryotes. IL-1α has pleiotropic effects on a wide range of cell types, and it has been extensively studied for its ability to contribute to various autoimmune and inflammation-linked disorders, including rheumatoid arthritis, Alzheimer's disease, systemic sclerosis and cardiovascular disorders. Interestingly, a significant proportion of IL-1α is translocated to the cell nucleus, in which it interacts with histone acetyltransferase complexes. Despite the importance of IL-1α, little is known regarding its binding targets and functions in the nucleus. We took advantage of the histone acetyltransferase (HAT) complexes being evolutionarily conserved from yeast to humans and the yeast SAGA complex serving as an epitome of the eukaryotic HAT complexes. Using gene knock-out technique and co-immunoprecipitation of the IL-1α precursor with TAP-tagged subunits of the yeast HAT complexes, we mapped the IL-1α-binding site to the HAT/Core module of the SAGA complex. We also predicted the 3-D structure of the IL-1α N-terminal domain, and by employing structure similarity searches, we found a similar structure in the C-terminal regulatory region of the catalytic subunit of the AMP-activated/Snf1 protein kinases, which interact with HAT complexes both in mammals and yeast, respectively. This finding is further supported with the ability of the IL-1α precursor to partially rescue growth defects of snf1Δ yeast strains on media containing 3-Amino-1,2,4-triazole (3-AT), a competitive inhibitor of His3. Finally, the careful evaluation of our data together with other published data in the field allows us to hypothesize a new function for the ADA complex in SAGA complex assembly.

Department of Genetics and Microbiology Faculty of Science Charles University Prague Prague Czech Republic

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Wade PA, Pruss D, Wolffe AP (1997) Histone acetylation: chromatin in action. Trends Biochem Sci 22: 128–132. PubMed

Allfrey VG, Faulkner R, Mirsky AE (1964) Acetylation and Methylation of Histones and Their Possible Role in the Regulation of Rna Synthesis. Proc Natl Acad Sci U S A 51: 786–794. PubMed PMC

Utley RT, Cote J (2003) The MYST family of histone acetyltransferases. Curr Top Microbiol Immunol 274: 203–236. PubMed

Brownell JE, Allis CD (1995) An activity gel assay detects a single, catalytically active histone acetyltransferase subunit in Tetrahymena macronuclei. Proc Natl Acad Sci U S A 92: 6364–6368. PubMed PMC

Brownell JE, Zhou J, Ranalli T, Kobayashi R, Edmondson DG, et al. (1996) Tetrahymena histone acetyltransferase A: a homolog to yeast Gcn5p linking histone acetylation to gene activation. Cell 84: 843–851. PubMed

Wang L, Mizzen C, Ying C, Candau R, Barlev N, et al. (1997) Histone acetyltransferase activity is conserved between yeast and human GCN5 and is required for complementation of growth and transcriptional activation. Mol Cell Biol 17: 519–527. PubMed PMC

Grant PA, Duggan L, Cote J, Roberts SM, Brownell JE, et al. (1997) Yeast Gcn5 functions in two multisubunit complexes to acetylate nucleosomal histones: characterization of an Ada complex and the SAGA (Spt/Ada) complex. Genes Dev 11: 1640–1650. PubMed

Wu PY, Ruhlmann C, Winston F, Schultz P (2004) Molecular architecture of the S. cerevisiae SAGA complex. Mol Cell 15: 199–208. PubMed

Lee KK, Workman JL (2007) Histone acetyltransferase complexes: one size doesn’t fit all. Nat Rev Mol Cell Biol 8: 284–295. PubMed

Lee KK, Sardiu ME, Swanson SK, Gilmore JM, Torok M, et al. (2011) Combinatorial depletion analysis to assemble the network architecture of the SAGA and ADA chromatin remodeling complexes. Mol Syst Biol 7: 503. PubMed PMC

Balasubramanian R, Pray-Grant MG, Selleck W, Grant PA, Tan S (2002) Role of the Ada2 and Ada3 transcriptional coactivators in histone acetylation. J Biol Chem 277: 7989–7995. PubMed

Horiuchi J, Silverman N, Marcus GA, Guarente L (1995) ADA3, a putative transcriptional adaptor, consists of two separable domains and interacts with ADA2 and GCN5 in a trimeric complex. Mol Cell Biol 15: 1203–1209. PubMed PMC

Horiuchi J, Silverman N, Pina B, Marcus GA, Guarente L (1997) ADA1, a novel component of the ADA/GCN5 complex, has broader effects than GCN5, ADA2, or ADA3. Mol Cell Biol 17: 3220–3228. PubMed PMC

Marcus GA, Horiuchi J, Silverman N, Guarente L (1996) ADA5/SPT20 links the ADA and SPT genes, which are involved in yeast transcription. Mol Cell Biol 16: 3197–3205. PubMed PMC

Dudley AM, Rougeulle C, Winston F (1999) The Spt components of SAGA facilitate TBP binding to a promoter at a post-activator-binding step in vivo. Genes Dev 13: 2940–2945. PubMed PMC

Bhaumik SR, Green MR (2002) Differential requirement of SAGA components for recruitment of TATA-box-binding protein to promoters in vivo. Mol Cell Biol 22: 7365–7371. PubMed PMC

Sermwittayawong D, Tan S (2006) SAGA binds TBP via its Spt8 subunit in competition with DNA: implications for TBP recruitment. Embo J 25: 3791–3800. PubMed PMC

Mohibullah N, Hahn S (2008) Site-specific cross-linking of TBP in vivo and in vitro reveals a direct functional interaction with the SAGA subunit Spt3. Genes Dev 22: 2994–3006. PubMed PMC

Grant PA, Schieltz D, Pray-Grant MG, Yates JR 3rd, Workman JL (1998) The ATM-related cofactor Tra1 is a component of the purified SAGA complex. Mol Cell 2: 863–867. PubMed

Saleh A, Schieltz D, Ting N, McMahon SB, Litchfield DW, et al. (1998) Tra1p is a component of the yeast Ada.Spt transcriptional regulatory complexes. J Biol Chem 273: 26559–26565. PubMed

Grant PA, Schieltz D, Pray-Grant MG, Steger DJ, Reese JC, et al. (1998) A subset of TAF(II)s are integral components of the SAGA complex required for nucleosome acetylation and transcriptional stimulation. Cell 94: 45–53. PubMed

Ingvarsdottir K, Krogan NJ, Emre NC, Wyce A, Thompson NJ, et al. (2005) H2B ubiquitin protease Ubp8 and Sgf11 constitute a discrete functional module within the Saccharomyces cerevisiae SAGA complex. Mol Cell Biol 25: 1162–1172. PubMed PMC

Kohler A, Pascual-Garcia P, Llopis A, Zapater M, Posas F, et al. (2006) The mRNA export factor Sus1 is involved in Spt/Ada/Gcn5 acetyltransferase-mediated H2B deubiquitinylation through its interaction with Ubp8 and Sgf11. Mol Biol Cell 17: 4228–4236. PubMed PMC

Sanders SL, Jennings J, Canutescu A, Link AJ, Weil PA (2002) Proteomics of the eukaryotic transcription machinery: identification of proteins associated with components of yeast TFIID by multidimensional mass spectrometry. Mol Cell Biol 22: 4723–4738. PubMed PMC

Pray-Grant MG, Daniel JA, Schieltz D, Yates JR 3rd, Grant PA (2005) Chd1 chromodomain links histone H3 methylation with SAGA- and SLIK-dependent acetylation. Nature 433: 434–438. PubMed

Eberharter A, Sterner DE, Schieltz D, Hassan A, Yates JR 3rd, et al (1999) The ADA complex is a distinct histone acetyltransferase complex in Saccharomyces cerevisiae. Mol Cell Biol 19: 6621–6631. PubMed PMC

Sterner DE, Grant PA, Roberts SM, Duggan LJ, Belotserkovskaya R, et al. (1999) Functional organization of the yeast SAGA complex: distinct components involved in structural integrity, nucleosome acetylation, and TATA-binding protein interaction. Mol Cell Biol 19: 86–98. PubMed PMC

Utley RT, Ikeda K, Grant PA, Cote J, Steger DJ, et al. (1998) Transcriptional activators direct histone acetyltransferase complexes to nucleosomes. Nature 394: 498–502. PubMed

Grant PA, Eberharter A, John S, Cook RG, Turner BM, et al. (1999) Expanded lysine acetylation specificity of Gcn5 in native complexes. J Biol Chem 274: 5895–5900. PubMed

Grant PA, Sterner DE, Duggan LJ, Workman JL, Berger SL (1998) The SAGA unfolds: convergence of transcription regulators in chromatin-modifying complexes. Trends Cell Biol 8: 193–197. PubMed

Dinarello CA (1996) Biologic basis for interleukin-1 in disease. Blood 87: 2095–2147. PubMed

Rainero I, Bo M, Ferrero M, Valfre W, Vaula G, et al. (2004) Association between the interleukin-1alpha gene and Alzheimer’s disease: a meta-analysis. Neurobiol Aging 25: 1293–1298. PubMed

Nicoll JA, Mrak RE, Graham DI, Stewart J, Wilcock G, et al. (2000) Association of interleukin-1 gene polymorphisms with Alzheimer’s disease. Ann Neurol 47: 365–368. PubMed PMC

Vicenova B, Vopalensky V, Burysek L, Pospisek M (2009) Emerging role of interleukin-1 in cardiovascular diseases. Physiol Res 58: 481–498. PubMed

Kawaguchi Y (1994) IL-1 alpha gene expression and protein production by fibroblasts from patients with systemic sclerosis. Clin Exp Immunol 97: 445–450. PubMed PMC

Wessendorf JH, Garfinkel S, Zhan X, Brown S, Maciag T (1993) Identification of a nuclear localization sequence within the structure of the human interleukin-1 alpha precursor. J Biol Chem 268: 22100–22104. PubMed

Yin H, Morioka H, Towle CA, Vidal M, Watanabe T, et al. (2001) Evidence that HAX-1 is an interleukin-1 alpha N-terminal binding protein. Cytokine 15: 122–137. PubMed

Hu B, Wang S, Zhang Y, Feghali CA, Dingman JR, et al. (2003) A nuclear target for interleukin-1alpha: interaction with the growth suppressor necdin modulates proliferation and collagen expression. Proc Natl Acad Sci U S A 100: 10008–10013. PubMed PMC

Pollock AS, Turck J, Lovett DH (2003) The prodomain of interleukin 1alpha interacts with elements of the RNA processing apparatus and induces apoptosis in malignant cells. Faseb J 17: 203–213. PubMed

Buryskova M, Pospisek M, Grothey A, Simmet T, Burysek L (2004) Intracellular interleukin-1alpha functionally interacts with histone acetyltransferase complexes. J Biol Chem 279: 4017–4026. PubMed

Ghaemmaghami S, Huh WK, Bower K, Howson RW, Belle A, et al. (2003) Global analysis of protein expression in yeast. Nature 425: 737–741. PubMed

Gueldener U, Heck S, Fielder T, Beinhauer J, Hegemann JH (1996) A new efficient gene disruption cassette for repeated use in budding yeast. Nucleic Acids Res 24: 2519–2524. PubMed PMC

Gietz RD, Woods RA (2002) Transformation of yeast by lithium acetate/single-stranded carrier DNA/polyethylene glycol method. Methods Enzymol 350: 87–96. PubMed

Werman A, Werman-Venkert R, White R, Lee JK, Werman B, et al. (2004) The precursor form of IL-1alpha is an intracrine proinflammatory activator of transcription. Proc Natl Acad Sci U S A 101: 2434–2439. PubMed PMC

Kim DE, Chivian D, Baker D (2004) Protein structure prediction and analysis using the Robetta server. Nucleic Acids Res 32: W526–531. PubMed PMC

Holm L, Rosenstrom P (2010) Dali server: conservation mapping in 3D. Nucleic Acids Res 38: W545–549. PubMed PMC

Collins SR, Kemmeren P, Zhao XC, Greenblatt JF, Spencer F, et al. (2007) Toward a comprehensive atlas of the physical interactome of Saccharomyces cerevisiae. Mol Cell Proteomics 6: 439–450. PubMed

Liu Y, Xu X, Singh-Rodriguez S, Zhao Y, Kuo MH (2005) Histone H3 Ser10 phosphorylation-independent function of Snf1 and Reg1 proteins rescues a gcn5- mutant in HIS3 expression. Mol Cell Biol 25: 10566–10579. PubMed PMC

Wilson MA, Koutelou E, Hirsch C, Akdemir K, Schibler A, et al. (2011) Ubp8 and SAGA regulate Snf1 AMP kinase activity. Mol Cell Biol 31: 3126–3135. PubMed PMC

Hardie DG (2007) AMP-activated/SNF1 protein kinases: conserved guardians of cellular energy. Nat Rev Mol Cell Biol 8: 774–785. PubMed

Gao G, Widmer J, Stapleton D, Teh T, Cox T, et al. (1995) Catalytic subunits of the porcine and rat 5′-AMP-activated protein kinase are members of the SNF1 protein kinase family. Biochim Biophys Acta 1266: 73–82. PubMed

Liu Y, Xu X, Kuo MH (2010) Snf1p regulates Gcn5p transcriptional activity by antagonizing Spt3p. Genetics 184: 91–105. PubMed PMC

Yousef AF, Xu GW, Mendez M, Brandl CJ, Mymryk JS (2008) Coactivator requirements for p53-dependent transcription in the yeast Saccharomyces cerevisiae. Int J Cancer 122: 942–946. PubMed

Smardova J, Smarda J, Koptikova J (2005) Functional analysis of p53 tumor suppressor in yeast. Differentiation 73: 261–277. PubMed

Flaman JM, Frebourg T, Moreau V, Charbonnier F, Martin C, et al. (1995) A simple p53 functional assay for screening cell lines, blood, and tumors. Proc Natl Acad Sci U S A 92: 3963–3967. PubMed PMC

Slovackova J, Grochova D, Navratilova J, Smarda J, Smardova J (2010) Transactivation by temperature-dependent p53 mutants in yeast and human cells. Cell Cycle 9: 2141–2148. PubMed

Orlova M, Kanter E, Krakovich D, Kuchin S (2006) Nitrogen availability and TOR regulate the Snf1 protein kinase in Saccharomyces cerevisiae. Eukaryot Cell 5: 1831–1837. PubMed PMC

Orlova M, Ozcetin H, Barrett L, Kuchin S (2010) Roles of the Snf1-activating kinases during nitrogen limitation and pseudohyphal differentiation in Saccharomyces cerevisiae. Eukaryot Cell 9: 208–214. PubMed PMC

Vincent O, Townley R, Kuchin S, Carlson M (2001) Subcellular localization of the Snf1 kinase is regulated by specific beta subunits and a novel glucose signaling mechanism. Genes Dev 15: 1104–1114. PubMed PMC

Nayak V, Zhao K, Wyce A, Schwartz MF, Lo WS, et al. (2006) Structure and dimerization of the kinase domain from yeast Snf1, a member of the Snf1/AMPK protein family. Structure 14: 477–485. PubMed

Momcilovic M, Hong SP, Carlson M (2006) Mammalian TAK1 activates Snf1 protein kinase in yeast and phosphorylates AMP-activated protein kinase in vitro. J Biol Chem 281: 25336–25343. PubMed

Towler MC, Hardie DG (2007) AMP-activated protein kinase in metabolic control and insulin signaling. Circ Res 100: 328–341. PubMed

Sterner DE, Belotserkovskaya R, Berger SL (2002) SALSA, a variant of yeast SAGA, contains truncated Spt7, which correlates with activated transcription. Proc Natl Acad Sci U S A 99: 11622–11627. PubMed PMC

Wu PY, Winston F (2002) Analysis of Spt7 function in the Saccharomyces cerevisiae SAGA coactivator complex. Mol Cell Biol 22: 5367–5379. PubMed PMC

Belotserkovskaya R, Sterner DE, Deng M, Sayre MH, Lieberman PM, et al. (2000) Inhibition of TATA-binding protein function by SAGA subunits Spt3 and Spt8 at Gcn4-activated promoters. Mol Cell Biol 20: 634–647. PubMed PMC

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