Cited by 450Genomics and Chromatin DynamicsFungal and yeast genetics researchNuclear Structure and Function

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A novel mediator between activator proteins and the RNA polymerase II transcription apparatus

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Kelleher, Raymond J., Flanagan, Peter M., Kornberg, Roger D.·1990·Cell

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Abstract

One gene activator protein may interfere with the effects of another in eukaryotic cells. We report here that a hybrid yeast-herpes gene activator protein inhibits transcriptional activation by a thymidine-rich DNA element in yeast. This example of activator interference can be faithfully reproduced in vitro. Interference is reversed by a partially purified yeast component, but not by RNA polymerase II or various polymerase II transcription factors. We conclude that the partially purified yeast component is a novel factor, and we suggest this factor mediates the transcriptional activation process.

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A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Bradford, M. M. (1976) · Analytical Biochemistry · doi:10.1016/0003-2697(76)90527-3 · Cited by 255,302 · cited 2×
“Protein was determined by the method of Bradford (1976) , using bovine serum albumin as a standard, and assays of P-galactosidase activity were performed as previously described (Miller, 1972; Guarente, 1983).”
Method
Transformation of intact yeast cells treated with alkali cations.
Ito, H., Fukuda, Y., Murata, K., & Kimura, A. (1984) · Journal of Bacteriology · doi:10.1271/bbb1961.48.341 · Cited by 4,235 · cited 1×
“Assays of BGaiactosidase Activity in Vivo Plasmid DNAs were introduced into Saccharomyces cerevisiae strain 5C (a his3-A200 ura3-52) by a lithium acetate transformation procedure (ito et al., 1983).”
Method
GAL4-VP16 is an unusually potent transcriptional activator
Sadowski, I., Ma, J., Triezenberg, S., & Ptashne, M. (1988) · Nature · doi:10.1038/335563a0 · Cited by 1,298 · cited 1×
“…GAL4-VP16, comprising the DNA binding domain (amino acids 1-147) of GAL4 fused to the activation domain (carboxy-terminal 78 amino acids) of VP16 (Sadowski et al., 1988), and a yeast factor whose binding site is a thymidine-rich sequence from upstream of the DED7 gene (T-rich binding factor; Lue…”
Method
Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast.
Guarente, L. P. (1983) · Methods in Enzymology · doi:10.1016/0076-6879(83)01013-7 · Cited by 1,022 · cited 1×
“Protein was determined by the method of Bradford (1976), using bovine serum albumin as a standard, and assays of P-galactosidase activity were performed as previously described (Miller, 1972; Guarente, 1983).”
Method
Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region.
Sawadogo, M., & Roeder, R. (1985) · Cell · doi:10.1016/0092-8674(85)90021-2 · Cited by 925 · cited 3×
“One line of evidence comes from alterations in the DNAase I footprint of partially purified HeLa TFIID on the adenovirus major late and E4 promoters caused by activator protein binding (Sawadogo and Roeder, 1985b; Horikoshi et al., 1988a, 1988b).”
Background
Functional dissection of VP16, the trans-activator of herpes simplex virus immediate early gene expression.Preprint
Triezenberg, S., Kingsbury, R., & McKnight, S. (1988) · Genes & Development · doi:10.1101/gad.2.6.718 · Cited by 785 · cited 1×
“For example, herpes viral protein 16 (VP16), an activator of immediate early gene expression in herpes virus-infected cells, causes a severalfold reduction in the level of transcription of an unrelated gene (Triezenberg et al., 1988).”
Background
A new class of yeast transcriptional activators.
Ma, J., & Ptashne, M. (1987) · Cell · doi:10.1016/0092-8674(87)90015-8 · Cited by 660 · cited 2×
“A single copy plasmid for expression of GAL4VP16 from the ADH7 plromoter, pCEN4:G4VPlG, was created by the insertion of the Kpnl fragment containing CEN4 sequences from pCZA into the unique Xbal site of pMA424 (gift of J. Ma and M. Ptashne; Ma and Ptashne, 1987) by blunt end ligation, leaving the 2pm origin intact but…”
Method
Negative effect of the transcriptional activator GAL4
Gill, G., & Ptashne, M. (1988) · Nature · doi:10.1038/334721a0 · Cited by 648 · cited 1×
“Similarly, high levels of the activator GAL4 in yeast cells diminish expression of genes lacking GAL4 binding sites as much as 5fold (Gill and Ptashne, 1988).”
Background
Steroid hormone receptors compete for factors that mediate their enhancer function.
Meyer, M., Gronemeyer, H., Turcotte, B., Bocquel, M., Tasset, D., & Chambon, P. (1989) · Cell · doi:10.1016/0092-8674(89)90918-5 · Cited by 542 · cited 2×
“In the case of the vertebrate estrogen, progesterone, and glucocorticoid receptors, which activate responsive genes in the presence of hormone, various pairwise combinations of both transfected and endogenous receptor genes exhibit interference in a hormone-dependent fashion (Meyer et al., 1989).”
Background
Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomeres in Saccharomyces cerevisiae
Buchman, A., Kimmerly, W. J., Rine, J., & Kornberg, R. (1988) · Molecular and Cellular Biology · doi:10.1128/mcb.8.1.210-225.1988 · Cited by 456 · cited 6×
“A similar inhibition of activation by the GRFI binding EN07 UAS was observed.”
BackgroundInfluentialSelf-citation

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Functional differences between yeast and human TFIID are localized to the highly conserved region.
Cormack, B., Strubin, M., Ponticelli, A., & Struhl, K. (1991) · Cell · doi:10.1016/0092-8674(91)90167-w · Cited by 101
“…by directly con- tatting TFIID or by interacting with a distinct protein that serves as an adaptor to the basic transcription machinery (Berger et al., 1990; Kambadur et al., 1990; Kelleher et al., 1990; Meisterernst et al., 1990; Pugh and Tjian, 1990) remains unanswered and controversial.”
Background
The multitalented Mediator complex
Carlsten, J. O. P., Zhu, X., & Gustafsson, C. M. (2013) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2013.08.007 · Cited by 92
Non-Recognition-of-BTH4, an Arabidopsis Mediator Subunit Homolog, Is Necessary for Development and Response to Salicylic Acid
Canet, J. V., Dobón, A., & Tornero, P. (2012) · The Plant Cell · doi:10.1105/tpc.112.103028 · Cited by 124
“(Kelleher et al., 1990; Flanagan et al., 1991).”
TFIID and human mediator coactivator complexes assemble cooperatively on promoter DNA.Preprint
Johnson, K. M., Wang, J., Smallwood, A., Arayata, C., & Carey, M. F. (2002) · Genes & Development · doi:10.1101/gad.995702 · Cited by 99
“The mediator coactivator complex was first purified from yeast as an activity that relieved squelching by the strong, model activator, GAL4-VP16 (Kelleher et al. 1990; Flanagan et al. 1991; Myers and Kornberg 2000).”
Background
Importance of Mediator complex in the regulation and integration of diverse signaling pathways in plants
Samanta, S., & Thakur, J. K. (2015) · Frontiers in Plant Science · doi:10.3389/fpls.2015.00757 · Cited by 107
“Mediator complex was first discovered in yeast in 1990, and within few years also reported in human (Kelleher et al., 1990; Flanagan et al., 1991; Thompson et al., 1993; Kim et al., 1994; Fondell et al., 1996; Ito et al., 1999).”
BackgroundInfluential
Transcriptional activation in an improved whole-cell extract from Saccharomyces cerevisiae
Woontner, M., Wade, P. A., Bonner, J. J., & Jaehning, J. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.9.4555-4560.1991 · Cited by 96
“The biochemical analysis of yeast transcription has progressed more slowly, in part because of the technical difficulties involved in preparing nuclei (14, 16).”
BackgroundMethod
N-ras oncogene causes AP-2 transcriptional self-interference, which leads to transformation.Preprint
Kannan, P., Buettner, R., Chiao, P., Yim, S., Sarkiss, M., & Tainsky, M. A. (1994) · Genes & Development · doi:10.1101/gad.8.11.1258 · Cited by 95
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Baniahmad, C., Nawaz, Z., Baniahmad, A., Gleeson, M., Tsai, M.-J., & O'Malley, B. (1995) · Molecular Endocrinology · doi:10.1210/mend.9.1.7760849 · Cited by 92
A Conserved Mediator Hinge Revealed in the Structure of the MED7·MED21 (Med7·Srb7) Heterodimer
Baumli, S., Hoeppner, S., & Cramer, P. (2005) · Journal of Biological Chemistry · doi:10.1074/jbc.m413466200 · Cited by 75
Yeast AP-1 like transcription factors (Yap) and stress response: a current overview
Rodrigues‐Pousada, C., Devaux, F., Caetano, S. M., Pimentel, C., da Silva, S. M., Gaspar‐Cordeiro, A., & Amaral, C. (2019) · Microbial Cell · doi:10.15698/mic2019.06.679 · Cited by 112
“The Mediator is a complex molecular machine composed of about 20 subunits organized in four domains (tail, middle, head and regulatory modules) [117-119].”
Background
Plant Mediator complex and its critical functions in transcription regulation
Yan, Y., Li, L., & Qu, L. (2015) · Journal of Integrative Plant Biology · doi:10.1111/jipb.12377 · Cited by 94
The plant Mediator complex and its role in jasmonate signaling
Zhai, Q., & Li, C. (2019) · Journal of Experimental Botany · doi:10.1093/jxb/erz233 · Cited by 101
Conformation of the RNA polymerase II C-terminal domain: circular dichroism of long and short fragments.
Bienkiewicz, E. A., Woody, A.-Y. M., & Woody, R. W. (2000) · Journal of Molecular Biology · doi:10.1006/jmbi.2000.3545 · Cited by 82
A major transactivator of varicella-zoster virus, the immediate-early protein IE62, contains a potent N-terminal activation domain
Perera, L. P., Mosca, J., Ruyechan, W., Hayward, G. S., Straus, S. E., & Hay, J. (1993) · Journal of Virology · doi:10.1128/jvi.67.8.4474-4483.1993 · Cited by 82
“This fragment, GAL4(1-147), which bears the N-terminal 147 amino acids, binds to DNA but does not activate transcription (6, 33).”
BackgroundMethodInfluential
The erythroid-specific protein cGATA-1 mediates distal enhancer activity through a specialized beta-globin TATA box.Preprint
Fong, T., & Emerson, B. M. (1992) · Genes & Development · doi:10.1101/gad.6.4.521 · Cited by 93
Transcriptional enhancers and their communication with gene promoters
Ray-Jones, H., & Spivakov, M. (2021) · Cellular and Molecular Life Sciences · doi:10.1007/s00018-021-03903-w · Cited by 81
A Genomic Screen in Yeast Reveals Novel Aspects of Nonstop mRNA Metabolism
Wilson, M. A., Meaux, S., & van Hoof, A. (2007) · Genetics · doi:10.1534/genetics.107.073205 · Cited by 78
“These genes have been implicated in both transcriptional activation and repression of a variety of genes, suggesting that they might increase his3-nonstop expression by increasing the strength of the HIS3 pro- moter (Kelleher et al. 1990; Flanagan et al. 1991; Thompson et al. 1993).”
Background
A highly conserved domain of TFIID displays species specificity in vivo.
Gill, G., & Tjian, R. (1991) · Cell · doi:10.1016/0092-8674(91)90166-v · Cited by 80
Accurate and efficient RNA polymerase II transcription with a soluble nuclear fraction derived from Drosophila embryos.
Kamakaka, R., Tyree, C. M., & Kadonaga, J. T. (1991) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.88.3.1024 · Cited by 80
CDK8 is associated with RAP2.6 and SnRK2.6 and positively modulates abscisic acid signaling and drought response in Arabidopsis
Zhu, Y., Huang, P., Guo, P., Chong, L., Yu, G., Sun, X., Hu, T., Li, Y., Hsu, C., Tang, K., et al. (2020) · New Phytologist · doi:10.1111/nph.16787 · Cited by 96
“Mediator is a large multi-protein complex that is evolutionarily conserved across eukaryotic species to serve as a co-regulator of gene transcription by connecting TFs to RNA polymerase II (RNAP II) (Kelleher et al., 1990; Myers & Kornberg, 2000; Carlsten et al., 2013).”
Background
Mediator Complex Recruits Epigenetic Regulators via Its Two Cyclin-dependent Kinase Subunits to Repress Transcription of Immune Response Genes
Tsutsui, T., Fukasawa, R., Shinmyouzu, K., Nakagawa, R., Tobe, K., Tanaka, A., & Ohkuma, Y. (2013) · Journal of Biological Chemistry · doi:10.1074/jbc.m113.486746 · Cited by 79
Mechanism of transcriptional antirepression by GAL4-VP16.Preprint
Croston, G. E., Laybourn, P. J., Paranjape, S. M., & Kadonaga, J. T. (1992) · Genes & Development · doi:10.1101/gad.6.12a.2270 · Cited by 91
“1991), and it appears that activation of transcription by GAL4-VP16 is dependent on an auxiliary activity that has been referred to as a mediator, adaptor, or intermediary factor (Berger et al. 1990; Kelleher et al. 1990; Flanagan et al. 1991; White et al. 1991).”
Background
trans-Activation of a globin promoter in nonerythroid cells.
Evans, T., & Felsenfeld, G. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.2.843 · Cited by 92
“One possibility is that Eryfl requires a coactivator, as has recently been suggested for activation by Spl and several viral transactivators (15, 19, 26); cell-specific coactivators might result in very different mechanisms of trans-activation, all still dependent on Eryfl.”
Background
Genes For Tfb2, Tfb3, and Tfb4 Subunits of Yeast Transcription/Repair Factor IIH
Feaver, W. J., Henry, N. L., Wang, Z.-G., Wu, X.-H., Svejstrup, J. Q., Bushnell, D., Friedberg, E., & Kornberg, R. (1997) · Journal of Biological Chemistry · doi:10.1074/jbc.272.31.19319 · Cited by 77
Self-citation
RNA polymerase II cofactor PC2 facilitates activation of transcription by GAL4-AH in vitro
Kretzschmar, M., Stelzer, G., Roeder, R., & Meisterernst, M. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.6.3927-3937.1994 · Cited by 75
The Oct-2 glutamine-rich and proline-rich activation domains can synergize with each other or duplicates of themselves to activate transcription
Tanaka, M., Clouston, W., & Herr, W. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.9.6046-6055.1994 · Cited by 74
“To prepare pCG-GAL4 (1-94)1XQ, pCG-GAL4(1-94)2XQ, pCG-GAL4(1-94)1XP, and pCG-GAL4(1-94)2xP, one or two tandem BbsI Q domain- or P domain-encoding fragments, as indicated, were inserted into the ACCA/TGGT BbsI cleavage site in pCGGAL4(1-94).”
BackgroundMethodInfluential
Transcriptional activation by the acidic domain of Vmw65 requires the integrity of the domain and involves additional determinants distinct from those necessary for TFIIB binding
Walker, S., Greaves, R., & O'Hare, P. (1993) · Molecular and Cellular Biology · doi:10.1128/mcb.13.9.5233-5244.1993 · Cited by 71
“Equally compelling evidence has accumulated for the involvement of an adaptor species, which is not necessary for basal transcription but is specifically required for the activation of transcription by the acidic domain (3, 15, 22, 30, 51, 57, 60).”
Background
A transcriptional mediator protein that is required for activation of many RNA polymerase II promoters and is conserved from yeast to humans
Lee, Y. C., Min, S., Gim, B., & Kim, Y.-J. (1997) · Molecular and Cellular Biology · doi:10.1128/mcb.17.8.4622 · Cited by 70
Identification, structure, and functional requirement of the Mediator submodule Med7N/31
Koschubs, T., Seizl, M., Larivière, L., Kurth, F., Baumli, S., Martin, D. E., & Cramer, P. (2008) · The EMBO Journal · doi:10.1038/emboj.2008.254 · Cited by 65
“Mediator was identified in yeast (Kelleher et al, 1990; Flanagan et al, 1991), has been purified from various fungi, metazoans, and plant (Bäckström et al, 2007), and was detected by homology analysis in many eukaryotic genomes (Boube et al, 2002; Bourbon, 2008).”
Caenorhabditis elegans mediator complexes are required for developmental-specific transcriptional activation.
Kwon, J., Park, J. M., Gim, B., Han, S.-J., Lee, J., & Kim, Y.-J. (1999) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.96.26.14990 · Cited by 68
Synergistic transcriptional activation by CTF/NF-I and the estrogen receptor involves stabilized interactions with a limiting target factor
Martinez, E., Dusserre, Y., Wahli, W., & Mermod, N. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.6.2937-2945.1991 · Cited by 68
“In these experiments, HeLa cells were transiently cotransfected with the cat reporter construct pG5BCAT containing five GAL4 binding sites upstream of the adenovirus Elb gene TATA box and cat sequences (27), together with either Gal 399-499 or the Gal VP16 activator, a GAL4(1-147) fusion with the negatively charged ac…”
Method
Evolution of disorder in Mediator complex and its functional relevance
Nagulapalli, M., Maji, S., Dwivedi, N., Dahiya, P., & Thakur, J. K. (2015) · Nucleic Acids Research · doi:10.1093/nar/gkv1135 · Cited by 87
Potentiation of glucocorticoid receptor-mediated gene expression by heat and chemical shock.
Sánchez, E., Hu, J., Zhong, S., Shen, P., Greene, M. J., & Housley, P. R. (1994) · Molecular Endocrinology · doi:10.1210/mend.8.4.8052262 · Cited by 66
Direct interaction of the tau 1 transactivation domain of the human glucocorticoid receptor with the basal transcriptional machinery
McEwan, I. J., Wright, A. P. H., Dahlman-Wright, K., Carlstedt-Duke, J., & Gustafsson, J.-Å. (1993) · Molecular and Cellular Biology · doi:10.1128/mcb.13.1.399-407.1993 · Cited by 65
Mediator: A key regulator of plant development
Buendía‐Monreal, M., & Gillmor, C. S. (2016) · Developmental Biology · doi:10.1016/j.ydbio.2016.06.009 · Cited by 78
Mediator of transcriptional regulation.
Björklund, S., & Kim, Y.-J. (1996) · TIBS -Trends in Biochemical Sciences. Regular ed · doi:10.1016/s0968-0004(96)10051-7 · Cited by 61
Cofactor squelching: Artifact or fact?
Schmidt, S. F., Larsen, B. D., Loft, A., & Mandrup, S. (2016) · BioEssays · doi:10.1002/bies.201600034 · Cited by 56
Conservation and evolution of transcriptional mechanisms in eukaryotes
Guarente, L., & Bermingham‐McDonogh, O. (1992) · Trends in Genetics · doi:10.1016/0168-9525(92)90021-u · Cited by 62
Twenty years of Mediator complex structural studies
Verger, A., Monté, D., & Villeret, V. (2019) · Biochemical Society Transactions · doi:10.1042/bst20180608 · Cited by 70
“The Kornberg laboratory isolated Mediator biochemically from a distinct crude yeast fraction that stimulated activator-dependent transcription in vitro using partially puri ﬁ ed proteins [1,2].”
Mediator – a universal complex in transcriptional regulation
Gustafsson, C., & Samuelsson, T. (2001) · Molecular Microbiology · doi:10.1046/j.1365-2958.2001.02481.x · Cited by 58
Cdk11-CyclinL Controls the Assembly of the RNA Polymerase II Mediator Complex
Drogat, J., Migeot, V., Mommaerts, E., Mullier, C., Dieu, M., van Bakel, H., & Hermand, D. (2012) · Cell Reports · doi:10.1016/j.celrep.2012.09.027 · Cited by 55
Tetracycline-regulated gene expression in the pathogen Ustilago maydis
Zarnack, K., Maurer, S. P., Kaffarnik, F., Ladendorf, O., Brachmann, A., Kämper, J., & Feldbrügge, M. (2006) · Fungal Genetics and Biology · doi:10.1016/j.fgb.2006.05.006 · Cited by 53
RNA polymerase II structure, and organization of the preinitiation complex.
Asturias, F. (2004) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2004.03.007 · Cited by 54
The Med1 subunit of the yeast mediator complex is involved in both transcriptional activation and repression.
Balčiūnas, D., Galman, C., Ronne, H., & Björklund, S. (1999) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.96.2.376 · Cited by 54
Mediator head subcomplex Med11/22 contains a common helix bundle building block with a specific function in transcription initiation complex stabilization
Seizl, M., Larivière, L., Pfaffeneder, T., Wenzeck, L., & Cramer, P. (2011) · Nucleic Acids Research · doi:10.1093/nar/gkr229 · Cited by 54
A Functional Module of Yeast Mediator That Governs the Dynamic Range of Heat-Shock Gene Expression
Singh, H., Erkine, A. M., Kremer, S. B., Duttweiler, H. M., Davis, D. A., Iqbal, J., Gross, R. R., & Gross, D. S. (2006) · Genetics · doi:10.1534/genetics.105.052738 · Cited by 47
Estradiol-inducible squelching and cell growth arrest by a chimeric VP16-estrogen receptor expressed in Saccharomyces cerevisiae: suppression by an allele of PDR1
Gilbert, D. M., Heery, D. M., Losson, R., Chambon, P., & Lemoine, Y. (1993) · Molecular and Cellular Biology · doi:10.1128/mcb.13.1.462-472.1993 · Cited by 53
“GAL4VP16 is an extremely toxic protein for yeast cells; successful transformation has been reported only when the protein is expressed from single-copy plasmids (21).”
Background
A Novel Drug-Regulated Gene Expression System Based on the Nuclear Receptor Constitutive Androstane Receptor (CAR)
Honkakoski, P., Jääskeläinen, I., Kortelahti, M., & Urtti, A. O. (2001) · Pharmaceutical Research · doi:10.1023/a:1011068015301 · Cited by 50
The Mechanism by Which the Human Apolipoprotein B Gene Reducer Operates Involves Blocking of Transcriptional Activation by Hepatocyte Nuclear Factor 3
Paulweber, B., Sandhofer, F., & Levy-Wilson, B. (1993) · Molecular and Cellular Biology · doi:10.1128/mcb.13.3.1534-1546.1993 · Cited by 52
“with the basic transcriptional machinery is currently under intense investigation, and evidence is accumulating that this signal transduction employs transcriptional mediator proteins (10, 11, 27, 36, 44).”
Background
Towards genome-wide prediction and characterization of enhancers in plants
Marand, A. P., Zhang, T., Zhu, B., & Jiang, J. (2016) · Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms · doi:10.1016/j.bbagrm.2016.06.006 · Cited by 53
“Upon DNA binding, TFs have been demonstrated to be responsible for recruiting the mediator complex, a major component of enhancerdirected transcription [10].”
Background
A kinase subunit of the human mediator complex, CDK8, positively regulates transcriptional activation
Furumoto, T., Tanaka, A., Ito, M., Malik, S., Hirose, Y., Hanaoka, F., & Ohkuma, Y. (2007) · Genes to Cells · doi:10.1111/j.1365-2443.2007.01036.x · Cited by 51
Transcription revisited: a commentary on the 1995 Cold Spring Harbor Laboratory meeting, "Mechanisms of Eukaryotic Transcription".Preprint
McKnight, S. L. (1996) · Genes & Development · doi:10.1101/gad.10.4.367 · Cited by 46
Mapping of a major surface-exposed site in herpes simplex virus protein Vmw65 to a region of direct interaction in a transcription complex assembly
Hayes, S., & O'Hare, P. (1993) · Journal of Virology · doi:10.1128/jvi.67.2.852-862.1993 · Cited by 50
“By direct analysis by affinity chromatography or transcription titration experiments, this region of Vmw65 has been shown to bind TFIID (59), TFIIB (36), and an adaptor species required for inducible gene expression but unnecessary for constitutive expression (5, 30, 39, 63).”
Background
Preparation and topology of the Mediator middle module
Koschubs, T., Lorenzen, K., Baumli, S., Sandström, S., Heck, A. J. R., & Cramer, P. (2010) · Nucleic Acids Research · doi:10.1093/nar/gkq029 · Cited by 50
“Med10 was also found to stably bind to Med14, confirming previous data (19) and showing that Med14 (1-259) is sufficient to establish a connection of middle module towards Mediator tail module.”
Result
Site-specific Srb10-dependent phosphorylation of the yeast Mediator subunit Med2 regulates gene expression from the 2-microm plasmid.
Hallberg, M., Polozkov, G. V., Hu, G.-Z., Béve, J., Gustafsson, C., Ronne, H., & Björklund, S. (2004) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.0400221101 · Cited by 49
Med9/Cse2 and Gal11 Modules Are Required for Transcriptional Repression of Distinct Group of Genes*
Han, S.-J., Lee, J., Kang, J., & Kim, Y.-J. (2001) · Journal of Biological Chemistry · doi:10.1074/jbc.m105596200 · Cited by 49
Modulating the potency of an activator in a yeast in vitro transcription system
Ohashi, Y., Brickman, J. M., Furman, E., Middleton, B., & Carey, M. F. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.4.2731-2739.1994 · Cited by 49
“The stimulatory response is a direct consequence of the VP16 activation domains because neither GAL4(1-147) nor GAL4(1-147)”
BackgroundMethodInfluential
Aromatic amino acid biosynthesis in the yeast Saccharomyces cerevisiae: a model system for the regulation of a eukaryotic biosynthetic pathway
Braus, G. (1991) · Microbiological Reviews · doi:10.1128/mr.55.3.349-370.1991 · Cited by 49
The Mediator complex of Caenorhabditis elegans: insights into the developmental and physiological roles of a conserved transcriptional coregulator
Grants, J. M., Goh, G. Y. S., & Taubert, S. (2015) · Nucleic Acids Research · doi:10.1093/nar/gkv037 · Cited by 50
The Chemistry of Regulation of Genes and Other Things
Ptashne, M. (2014) · Journal of Biological Chemistry · doi:10.1074/jbc.x114.547323 · Cited by 47
“, the mediator) was used as an early assay to isolate the mediator (70).”
Method
Yeast and human TFIIDs are interchangeable for the response to acidic transcriptional activators in vitro.
Kelleher, R., Flanagan, P., Chasman, D. I., Ponticelli, A., Struhl, K., & Kornberg, R. (1992) · Genes & Development · doi:10.1101/gad.6.2.296 · Cited by 46
Self-citation
The interactions of transcription factors and their adaptors, coactivators and accessory proteins
Martin, K. (1991) · Bioessays · doi:10.1002/bies.950131003 · Cited by 46
The Mediator complex subunit 8 regulates organ size in Arabidopsis thaliana
Xu, R., & Li, Y.-H. (2012) · Plant Signalling & Behavior · doi:10.4161/psb.18803 · Cited by 45
Transcriptomic correlates of electrophysiological and morphological diversity within and across excitatory and inhibitory neuron classes
Bomkamp, C., Tripathy, S. J., Gonzales, C. B., Hjerling‐Leffler, J., Craig, A. M., & Pavlidis, P. (2019) · PLoS Computational Biology · doi:10.1371/journal.pcbi.1007113 · Cited by 42
Mediator subunit MED31 is required for radial patterning of Arabidopsis roots
Zhang, X., Zhou, W., Chen, Q., Fang, M., Zheng, S., Scheres, B., & Li, C. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1800592115 · Cited by 35
“Since its identification in yeast (32, 33) and animals (34), Mediator has been extensively investigated for its ability to orchestrate the transcription factor-dependent assembly of the Pol II preinitiation complex (PIC) via discrete interactions with signal-dependent transcription factors and Pol II (26–29).”
Background
Interactions between subunits of Drosophila Mediator and activator proteins.
Kim, Y.-J., & Lis, J. (2005) · TIBS -Trends in Biochemical Sciences. Regular ed · doi:10.1016/j.tibs.2005.03.010 · Cited by 44
The Coactivator p/CIP/SRC-3 Facilitates Retinoic Acid Receptor Signaling via Recruitment of GCN5*
Brown, K., Chen, Y., Underhill, T., Mymryk, J., & Torchia, J. (2003) · Journal of Biological Chemistry · doi:10.1074/jbc.m307832200 · Cited by 44
Emerging roles of Cdk8 in cell cycle control
Szilágyi, Z., & Gustafsson, C. M. (2013) · Biochimica et Biophysica Acta (BBA) - Gene Regulatory Mechanisms · doi:10.1016/j.bbagrm.2013.04.010 · Cited by 46
“transcription in vitro [1,2].”
Biochemical and genetic characterization of a yeast TFIID mutant that alters transcription in vivo and DNA binding in vitro
Arndt, K. M., Ricupero, S. L., Eisenmann, D., & Winston, F. (1992) · Molecular and Cellular Biology · doi:10.1128/mcb.12.5.2372-2382.1992 · Cited by 43
Inhibition of Chromatin Assembly in Xenopus Oocytes Correlates with Derepression of the Mouse Mammary Tumor Virus Promoter
Perlmann, T., & Wränge, Ö. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.10.5259-5265.1991 · Cited by 46
“1 ng per oocyte) factors of the transcription machinery, are evidently utilized rV wt, 5 ng per oocyte) as in transcriptional regulation (2, 14, 15, 25).”
Background
Specific functions for Mediator complex subunits from different modules in the transcriptional response of Arabidopsis thaliana to abiotic stress
Crawford, T. S., Karamat, F., Lehotai, N., Rentoft, M., Blomberg, J., Strand, Å., & Björklund, S. (2020) · Scientific Reports · doi:10.1038/s41598-020-61758-w · Cited by 46
Mediator Phosphorylation Prevents Stress Response Transcription During Non-stress Conditions
Miller, C., Matić, I., Maier, K. C., Schwalb, B., Röther, S., Sträßer, K., Tresch, A., Mann, M., & Cramer, P. (2012) · Journal of Biological Chemistry · doi:10.1074/jbc.m112.430140 · Cited by 44
Transcriptional activation by herpes simplex virus type 1 VP16 in vitro and its inhibition by oligopeptides
Wu, T.-J., Monokian, G., Mark, D., & Wobbe, C. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.5.3484-3493.1994 · Cited by 42
“Numerous examples of this latter phenomenon, often referred to as squelching, have been observed in vivo and in vitro and are thought to result from the sequestration of a limiting factor necessary for activated transcription (4, 21, 30, 42).”
Background
Salt stress activates the CDK8-AHL10-SUVH2/9 module to dynamically regulate salt tolerance in Arabidopsis
Guo, P., Chong, L., Jiao, Z., Xu, R., Niu, Q., & Zhu, Y. (2025) · Nature Communications · doi:10.1038/s41467-025-57806-6 · Cited by 42
Protein Dimerization Generates Bistability in Positive Feedback Loops
Hsu, C., Jaquet, V., Gençoğlu, M., & Becskei, A. (2016) · Cell Reports · doi:10.1016/j.celrep.2016.06.072 · Cited by 34
Initiation on chromatin templates in a yeast RNA polymerase II transcription system.Preprint
Lorch, Y., LaPointe, J. W., & Kornberg, R. (1992) · Genes & Development · doi:10.1101/gad.6.12a.2282 · Cited by 41
Self-citation
The conserved carboxy-terminal domain of Saccharomyces cerevisiae TFIID is sufficient to support normal cell growth
Poon, D., Schroeder, S., Wang, C.-K., Yamamoto, T., Horikoshi, M., Roeder, R., & Weil, P. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.10.4809-4821.1991 · Cited by 41
Differential ability of proximal and remote element pairs to cooperate in activating RNA polymerase II transcription
Wang, W.-D., & Gralla, J. (1991) · Molecular and Cellular Biology · doi:10.1128/mcb.11.9.4561-4571.1991 · Cited by 41
“of cooperation between upstream and proximal elements is now receiving more attention since recent studies have indicated that there may be adapter factors that mediate the interactions between the upstream and downstream elements (4, 20, 29).”
Background
Two conserved modules of Schizosaccharomyces pombe Mediator regulate distinct cellular pathways
Linder, T., Rasmussen, N. N., Samuelsen, C. O., Chatzidaki, E., Baraznenok, V., Béve, J., Henriksen, P., Gustafsson, C., & Holmberg, S. (2008) · Nucleic Acids Research · doi:10.1093/nar/gkn070 · Cited by 40
High level expression of the major transactivation domain of the human glucocorticoid receptor in yeast cells inhibits endogenous gene expression and cell growth.
Wright, A. P. H., McEwan, I. J., Dahlman-Wright, K., & Gustafsson, J.-Å. (1991) · Molecular Endocrinology · doi:10.1210/mend-5-10-1366 · Cited by 40
“In vitro squelching data suggest that VP16 interacts with proteins in the transcriptional initiation complex indirectly via an adaptor protein (23, 24).”
Background
Med1 Subunit of the Mediator Complex in Nuclear Receptor-Regulated Energy Metabolism, Liver Regeneration, and Hepatocarcinogenesis
Jia, Y., Viswakarma, N., & Reddy, J. K. (2014) · Gene Expression · doi:10.3727/105221614x13919976902219 · Cited by 54
Human mediator kinase subunit CDK11 plays a negative role in viral activator VP16‐dependent transcriptional regulation
Tsutsui, T., Umemura, H., Tanaka, A., Mizuki, F., Hirose, Y., & Ohkuma, Y. (2008) · Genes to Cells · doi:10.1111/j.1365-2443.2008.01208.x · Cited by 38
The molecular basis of eucaryotic transcription
Kornberg, R. (2007) · Cell Death and Differentiation · doi:10.1038/sj.cdd.4402251 · Cited by 38
Self-citation
The classical srb4-138 mutant allele causes dissociation of yeast Mediator.
Linder, T., Zhu, X., Baraznenok, V., & Gustafsson, C. (2006) · Biochemical and Biophysical Research Communications - BBRC · doi:10.1016/j.bbrc.2006.08.099 · Cited by 38
Identification of functional targets of the Zta transcriptional activator by formation of stable preinitiation complex intermediates
Lieberman, P. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.12.8365-8375.1994 · Cited by 37
“from the basal-level transcription factors (3, 26, 27).”
Background
MED15, encoding a subunit of the mediator complex, is overexpressed at high frequency in castration‐resistant prostate cancer
Shaikhibrahim, Z., Menon, R., Braun, M., Offermann, A., Queisser, A., Boehm, D., Vogel, W., Rüenauver, K., Ruiz, C., Zellweger, T., et al. (2014) · International Journal of Cancer · doi:10.1002/ijc.28647 · Cited by 36
Genetic evidence for the interaction of the yeast transcriptional co-activator proteins GCN5 and ADA2
Georgakopoulos, T., Gounalaki, N., & Thireos, G. (1995) · Molecular and General Genetics MGG · doi:10.1007/bf00290718 · Cited by 36
“The existence of such factors, termed mediators or adaptors, which confer the ability to respond to activator proteins in reconstituted systems, has been shown in both yeast and mammalian systems ( Kelleher et al. 1990; Flanagan et al. 1991; Kretzschmer et al. 1993).”
Background
Activation domains of transcriptional regulatory proteins
Johnson, P. F., Sterneck, E., & Williams, S. C. (1993) · The Journal of Nutritional Biochemistry · doi:10.1016/0955-2863(93)90069-9 · Cited by 33
Mechanistic advances in eukaryotic gene activation.
Carey, M. F. (1991) · Current Opinion in Cell Biology · doi:10.1016/0955-0674(91)90073-8 · Cited by 36
Molecular architecture of enhancer–promoter interaction
Hamamoto, K., & Fukaya, T. (2022) · Current Opinion in Cell Biology · doi:10.1016/j.ceb.2022.01.003 · Cited by 58
Transcriptional Silencing of MCL-1 Through Cyclin-Dependent Kinase Inhibition in Acute Myeloid Leukemia
Tibes, R., & Bogenberger, J. M. (2019) · Frontiers in Oncology · doi:10.3389/fonc.2019.01205 · Cited by 42
Emerging functions of multi-protein complex Mediator with special emphasis on plants
Malik, N., Agarwal, P., & Tyagi, A. K. (2017) · Critical Reviews in Biochemistry and Molecular Biology · doi:10.1080/10409238.2017.1325830 · Cited by 41
“It was first reported in yeast (Kelleher et al. 1990, Flanagan et al. 1991), followed by its isolation in mouse (Jiang et al. 1998), and thereafter, its presence in other eukaryotes including plants was established through biochemical purification and comparative genomic studies (Gu et al. 1999,…”
Background
Species-Specific Interaction of the Glutamine-Rich Activation Domains of Spl with the TATA Box-Binding Protein
Emili, A., Greenblatt, J., & Ingles, C. J. (1994) · Molecular and Cellular Biology · doi:10.1128/mcb.14.3.1582-1593.1994 · Cited by 63
“By several genetic and biochemical criteria, acidic activators also depend on cofactors for function (2, 14, 30, 56, 57).”
Background
Evolution of sequence repetition and gene duplications in the TATA-binding protein TBP (TFIID).
Hancock, J. M. (1993) · Nucleic Acids Research · doi:10.1093/nar/21.12.2823 · Cited by 35
Characterization of cis-acting elements regulating transcription from the promoter of a constitutively active rice actin gene
Wang, Y., Zhang, W., Cao, J., Mcelroy, D., & Wu, R. (1992) · Molecular and Cellular Biology · doi:10.1128/mcb.12.8.3399-3406.1992 · Cited by 35
“It stimulates transcription from templates containing poly(dA-dT) elements as much as 30-fold (24, 28).”
Background
Transcriptional activation by the pseudorabies virus immediate early protein.Preprint
Martin, K., Lillie, J., & Green, M. (1990) · Genes & Development · doi:10.1101/gad.4.12b.2376 · Cited by 35
“Squelching experiments have been used to compare the targets of various transcriptional activators (Meyer et al. 1989; Berger et al. 1990; Kelleher et al. 1990; Martin et al. 1990).”
Background
A Functional Portrait of Med7 and the Mediator Complex in Candida albicans
Tebbji, F., Chen, Y., Albert, J. R., Gunsalus, K. T. W., Kumamoto, C. A., Nantel, A., Sellam, A., & Whiteway, M. S. (2014) · PLoS Genetics · doi:10.1371/journal.pgen.1004770 · Cited by 34
PEA1 and PEA3 enhancer elements are primary components of the polyomavirus late transcription initiator element
Yoo, W., Martin, M. E., & Folk, W. R. (1991) · Journal of Virology · doi:10.1128/jvi.65.10.5391-5400.1991 · Cited by 34
Concerted stimulation of transcription by glucocorticoid receptors and basal transcription factors: limited transcriptional synergism suggests mediation by coactivators/adaptors.
Bastian, L. S., & Nordeen, S. (1991) · Molecular Endocrinology · doi:10.1210/mend-5-5-619 · Cited by 34
“An alternative model to cooperative DNA binding has been proposed which suggests that transcription factors synergistically activate the RNA polymerase-ll complex by interacting through intermediary factors that directly or indirectly transfer the activation signal to the transcription initiation complex (1, 2, 10-14).”
Background
The tail-module of yeast Mediator complex is required for telomere heterochromatin maintenance
Peng, J., & Zhou, J. (2011) · Nucleic Acids Research · doi:10.1093/nar/gkr757 · Cited by 34

Authors: Kelleher, R. J., Flanagan, P. M., & Kornberg, R. D.
Affiliations: Stanford Medicine (US)
ORCID: Kornberg, Roger D.
Venue: Cell
Publisher: Elsevier BV
Published: 1990-06
Volume: 61
Issue: 7
Pages: 1209-1215
ISSN: 0092-8674 (ISSN-L)
DOI: 10.1016/0092-8674(90)90685-8
OpenAlex: W2021893003
Cited by: 450
References: 35
Citations by year: 2026: 5 · 2025: 6 · 2024: 10 · 2023: 3 · 2022: 5 · 2021: 13 · 2020: 20 · 2019: 12 · 2018: 14 · 2017: 10 · 2016: 8 · 2015: 8 · 2014: 10 · 2013: 16 · 2012: 8
FWCI: 20.26
Type: journal article
Language: en
Open access: hybrid
doi.org
Licence: cc-by-nc-nd
www.elsevier.com/tdm/userlicense/1.0/ (tdm)
www.elsevier.com/legal/tdmrep-license (tdm)
creativecommons.org/licenses/by-nc-nd/4.0/ (vor, 5436 d)
Locations: Cell (publishedVersion, cc-by-nc-nd)
PubMed (publishedVersion)
Funders: National Institutes of Health (GM-07365, GM-36859)
Topics: Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Nuclear Structure and Function
Keywords: RNA polymerase II, transcriptional activation, transcriptional coactivator, transcription factors
MeSH terms: beta-Galactosidase, Cell Nucleus, DNA-Binding Proteins, DNA, Fungal, Fungal Proteins, Kinetics, Phosphoproteins, Plasmids, Recombinant Fusion Proteins, RNA Polymerase II, Saccharomyces cerevisiae, Transcription Factors, Transcription, Genetic, Trans-Activators, Gene Expression Regulation, Fungal, Simplexvirus, Saccharomyces cerevisiae Proteins

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A novel mediator between activator proteins and the RNA polymerase II transcription apparatus

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