Note to users. If you're seeing this message, it means that your browser cannot find this page's style/presentation instructions -- or possibly that you are using a browser that does not support current Web standards. Find out more about why this message is appearing, and what you can do to make your experience of our site the best it can be.
Structure, Function, and Activator-Induced Conformations of the CRSP Coactivator
Dylan J. Taatjes,1Anders M. Näär,1Frank Andel III,12Eva Nogales,12Robert Tjian1*
The human cofactor complexes ARC (activator-recruited cofactor) and
CRSP (cofactor required for Sp1 activation) mediate activator-dependenttranscription in vitro. Although these complexes share severalcommon
subunits, their structural and functional relationshipsremain unknown.
Here, we report that affinity-purified ARC consistsof two distinct
multisubunit complexes: a larger complex, denotedARC-L, and a smaller
coactivator, CRSP. Reconstituted in vitrotranscription with
biochemically separated ARC-L and CRSP revealsdifferential cofactor
functions. The ARC-L complex is transcriptionallyinactive, whereas the
CRSP complex is highly active. Structuraldetermination by electron
microscopy (EM) and three-dimensionalreconstruction indicate
substantial differences in size and shapebetween ARC-L and CRSP.
Moreover, EM analysis of independentlyderived CRSP complexes reveals
distinct conformations inducedby different activators. These results
suggest that CRSP may potentiatetranscription via specific
activator-induced conformational changes.
1 Howard Hughes Medical Institute and
2 Lawrence Berkeley National Laboratory, Department
of Molecular and Cell Biology, 401 Barker Hall, University of
California, Berkeley, CA 94720, USA.
*
To whom correspondence should be addressed. E-mail:
jmlim{at}uclink4.berkeley.edu
The editors suggest the following Related Resources on Science sites:
In Science Magazine
PERSPECTIVES
Michael Meisterernst (8 February 2002) Science295 (5557), 984.
[DOI: 10.1126/science.1069485] |Summary »|Full Text »|PDF »
Transcription initiation by human RNA polymerase II visualized at single-molecule resolution.
A. Revyakin, Z. Zhang, R. A. Coleman, Y. Li, C. Inouye, J. K. Lucas, S.-R. Park, S. Chu, and R. Tjian (2012)
Genes & Dev.
26, 1691-1702
|Abstract »|Full Text »|PDF »
Core promoter-selective function of HMGA1 and Mediator in Initiator-dependent transcription.
M. Xu, P. Sharma, S. Pan, S. Malik, R. G. Roeder, and E. Martinez (2011)
Genes & Dev.
25, 2513-2524
|Abstract »|Full Text »|PDF »
The Mediator Complex in Plants: Structure, Phylogeny, and Expression Profiling of Representative Genes in a Dicot (Arabidopsis) and a Monocot (Rice) during Reproduction and Abiotic Stress.
S. Mathur, S. Vyas, S. Kapoor, and A. K. Tyagi (2011)
Plant Physiology
157, 1609-1627
|Abstract »|Full Text »|PDF »
Transcriptional Regulation in Saccharomyces cerevisiae: Transcription Factor Regulation and Function, Mechanisms of Initiation, and Roles of Activators and Coactivators.
MED12, the Mediator Complex Subunit 12 Gene, Is Mutated at High Frequency in Uterine Leiomyomas.
N. Makinen, M. Mehine, J. Tolvanen, E. Kaasinen, Y. Li, H. J. Lehtonen, M. Gentile, J. Yan, M. Enge, M. Taipale, et al. (2011)
Science
334, 252-255
|Abstract »|Full Text »|PDF »
Herpes Simplex Virus 1 ICP4 Forms Complexes with TFIID and Mediator in Virus-Infected Cells.
Understanding large multiprotein complexes: applying a multiple allosteric networks model to explain the function of the Mediator transcription complex.
Structures of three distinct activator-TFIID complexes.
W.-L. Liu, R. A. Coleman, E. Ma, P. Grob, J. L. Yang, Y. Zhang, G. Dailey, E. Nogales, and R. Tjian (2009)
Genes & Dev.
23, 1510-1521
|Abstract »|Full Text »|PDF »
The human CDK8 subcomplex is a molecular switch that controls Mediator coactivator function.
M. T. Knuesel, K. D. Meyer, C. Bernecky, and D. J. Taatjes (2009)
Genes & Dev.
23, 439-451
|Abstract »|Full Text »|PDF »
The Human CDK8 Subcomplex Is a Histone Kinase That Requires Med12 for Activity and Can Function Independently of Mediator.
M. T. Knuesel, K. D. Meyer, A. J. Donner, J. M. Espinosa, and D. J. Taatjes (2009)
Mol. Cell. Biol.
29, 650-661
|Abstract »|Full Text »|PDF »
MED19 and MED26 Are Synergistic Functional Targets of the RE1 Silencing Transcription Factor in Epigenetic Silencing of Neuronal Gene Expression.
N. Ding, C. Tomomori-Sato, S. Sato, R. C. Conaway, J. W. Conaway, and T. G. Boyer (2009)
J. Biol. Chem.
284, 2648-2656
|Abstract »|Full Text »|PDF »
A Key Transcription Cofactor on the Nascent Sex Chromosomes of European Tree Frogs (Hyla arborea).
H. Niculita-Hirzel, M. Stock, and N. Perrin (2008)
Genetics
179, 1721-1723
|Abstract »|Full Text »|PDF »
MED1 Phosphorylation Promotes Its Association with Mediator: Implications for Nuclear Receptor Signaling.
M. Belakavadi, P. K. Pandey, R. Vijayvargia, and J. D. Fondell (2008)
Mol. Cell. Biol.
28, 3932-3942
|Abstract »|Full Text »|PDF »
Pygopus activates Wingless target gene transcription through the mediator complex subunits Med12 and Med13.
I. Carrera, F. Janody, N. Leeds, F. Duveau, and J. E. Treisman (2008)
PNAS
105, 6644-6649
|Abstract »|Full Text »|PDF »
Phosphorylation by c-Jun NH2-terminal Kinase 1 Regulates the Stability of Transcription Factor Sp1 during Mitosis.
Cdk8 Is Essential for Preimplantation Mouse Development.
T. Westerling, E. Kuuluvainen, and T. P. Makela (2007)
Mol. Cell. Biol.
27, 6177-6182
|Abstract »|Full Text »|PDF »
A Genetic Screen Identifies Novel Polycomb Group Genes in Drosophila.
A. G. de Ayala Alonso, L. Gutierrez, C. Fritsch, B. Papp, D. Beuchle, and J. Muller (2007)
Genetics
176, 2099-2108
|Abstract »|Full Text »|PDF »
The VP16 Activation Domain Establishes an Active Mediator Lacking CDK8 in Vivo.
T. Uhlmann, S. Boeing, M. Lehmbacher, and M. Meisterernst (2007)
J. Biol. Chem.
282, 2163-2173
|Abstract »|Full Text »|PDF »
Yeast TFIID Serves as a Coactivator for Rap1p by Direct Protein-Protein Interaction.
K. A. Garbett, M. K. Tripathi, B. Cencki, J. H. Layer, and P. A. Weil (2007)
Mol. Cell. Biol.
27, 297-311
|Abstract »|Full Text »|PDF »
Quantitative proteomic analysis of distinct mammalian Mediator complexes using normalized spectral abundance factors.
A. C. Paoletti, T. J. Parmely, C. Tomomori-Sato, S. Sato, D. Zhu, R. C. Conaway, J. W. Conaway, L. Florens, and M. P. Washburn (2006)
PNAS
103, 18928-18933
|Abstract »|Full Text »|PDF »
Mediator Modulates Gli3-Dependent Sonic Hedgehog Signaling.
H. Zhou, S. Kim, S. Ishii, and T. G. Boyer (2006)
Mol. Cell. Biol.
26, 8667-8682
|Abstract »|Full Text »|PDF »
The cyclin-dependent kinase 8 module sterically blocks Mediator interactions with RNA polymerase II.
H. Elmlund, V. Baraznenok, M. Lindahl, C. O. Samuelsen, P. J. B. Koeck, S. Holmberg, H. Hebert, and C. M. Gustafsson (2006)
PNAS
103, 15788-15793
|Abstract »|Full Text »|PDF »
Human Mediator Enhances Basal Transcription by Facilitating Recruitment of Transcription Factor IIB during Preinitiation Complex Assembly.
H. J. Baek, Y. K. Kang, and R. G. Roeder (2006)
J. Biol. Chem.
281, 15172-15181
|Abstract »|Full Text »|PDF »
Mediator Is a Transducer of Wnt/beta-Catenin Signaling.
S. Kim, X. Xu, A. Hecht, and T. G. Boyer (2006)
J. Biol. Chem.
281, 14066-14075
|Abstract »|Full Text »|PDF »
Hsp90{alpha} Recruited by Sp1 Is Important for Transcription of 12(S)-Lipoxygenase in A431 Cells.
J.-J. Hung, C.-Y. Wu, P.-C. Liao, and W.-C. Chang (2005)
J. Biol. Chem.
280, 36283-36292
|Abstract »|Full Text »|PDF »
A structural perspective of CTD function.
A. Meinhart, T. Kamenski, S. Hoeppner, S. Baumli, and P. Cramer (2005)
Genes & Dev.
19, 1401-1415
|Abstract »|Full Text »|PDF »
A Conserved Mediator Hinge Revealed in the Structure of the MED7{middle dot}MED21 (Med7{middle dot}Srb7) Heterodimer.
S. Baumli, S. Hoeppner, and P. Cramer (2005)
J. Biol. Chem.
280, 18171-18178
|Abstract »|Full Text »|PDF »
Components of the transcriptional Mediator complex are required for asymmetric cell division in C. elegans.
A. Yoda, H. Kouike, H. Okano, and H. Sawa (2005)
Development
132, 1885-1893
|Abstract »|Full Text »|PDF »
Structural and Functional Characterization of PC2 and RNA Polymerase II-Associated Subpopulations of Metazoan Mediator.
S. Malik, H. J. Baek, W. Wu, and R. G. Roeder (2005)
Mol. Cell. Biol.
25, 2117-2129
|Abstract »|Full Text »|PDF »
A high resolution protein interaction map of the yeast Mediator complex.
B. Guglielmi, N. L. van Berkum, B. Klapholz, T. Bijma, M. Boube, C. Boschiero, H.-M. Bourbon, F. C. P. Holstege, and M. Werner (2004)
Nucleic Acids Res.
32, 5379-5391
|Abstract »|Full Text »|PDF »
Structural and Functional Organization of TRAP220, the TRAP/Mediator Subunit That Is Targeted by Nuclear Receptors.
S. Malik, M. Guermah, C.-X. Yuan, W. Wu, S. Yamamura, and R. G. Roeder (2004)
Mol. Cell. Biol.
24, 8244-8254
|Abstract »|Full Text »|PDF »
MED16 and MED23 of Mediator are coactivators of lipopolysaccharide- and heat-shock-induced transcriptional activators.
T. W. Kim, Y.-J. Kwon, J. M. Kim, Y.-H. Song, S. N. Kim, and Y.-J. Kim (2004)
PNAS
101, 12153-12158
|Abstract »|Full Text »|PDF »
Transcriptional Coactivator PC4 Stimulates Promoter Escape and Facilitates Transcriptional Synergy by GAL4-VP16.
A. Fukuda, T. Nakadai, M. Shimada, T. Tsukui, M. Matsumoto, Y. Nogi, M. Meisterernst, and K. Hisatake (2004)
Mol. Cell. Biol.
24, 6525-6535
|Abstract »|Full Text »|PDF »
The Caenorhabditis elegans Ortholog of TRAP240, CeTRAP240/let-19, Selectively Modulates Gene Expression and Is Essential for Embryogenesis.
J.-C. Wang, A. Walker, T. K. Blackwell, and K. R. Yamamoto (2004)
J. Biol. Chem.
279, 29270-29277
|Abstract »|Full Text »|PDF »
The activator-recruited cofactor/Mediator coactivator subunit ARC92 is a functionally important target of the VP16 transcriptional activator.
F. Yang, R. DeBeaumont, S. Zhou, and A. M. Naar (2004)
PNAS
101, 2339-2344
|Abstract »|Full Text »|PDF »
RNA Polymerase II (Pol II)-TFIIF and Pol II-Mediator Complexes: the Major Stable Pol II Complexes and Their Activity in Transcription Initiation and Reinitiation.
P. G. Rani, J. A. Ranish, and S. Hahn (2004)
Mol. Cell. Biol.
24, 1709-1720
|Abstract »|Full Text »|PDF »
The mediator coactivator complex: functional and physical roles in transcriptional regulation.
Human Mediator Enhances Activator-Facilitated Recruitment of RNA Polymerase II and Promoter Recognition by TATA-Binding Protein (TBP) Independently of TBP-Associated Factors.
Two subunits of the Drosophila mediator complex act together to control cell affinity.
F. Janody, Z. Martirosyan, A. Benlali, and J. E. Treisman (2003)
Development
130, 3691-3701
|Abstract »|Full Text »|PDF »
TRAP230/ARC240 and TRAP240/ARC250 Mediator subunits are functionally conserved through evolution.
C. O. Samuelsen, V. Baraznenok, O. Khorosjutina, H. Spahr, T. Kieselbach, S. Holmberg, and C. M. Gustafsson (2003)
PNAS
100, 6422-6427
|Abstract »|Full Text »|PDF »
Transactivation of the Progesterone Receptor Gene in Granulosa Cells: Evidence that Sp1/Sp3 Binding Sites in the Proximal Promoter Play a Key Role in Luteinizing Hormone Inducibility.
V. Sriraman, S. C. Sharma, and J. S. Richards (2003)
Mol. Endocrinol.
17, 436-449
|Abstract »|Full Text »|PDF »
Mediator and p300/CBP-Steroid Receptor Coactivator Complexes Have Distinct Roles, but Function Synergistically, during Estrogen Receptor {alpha}-Dependent Transcription with Chromatin Templates.
A Complex of the Srb8, -9, -10, and -11 Transcriptional Regulatory Proteins from Yeast.
T. Borggrefe, R. Davis, H. Erdjument-Bromage, P. Tempst, and R. D. Kornberg (2002)
J. Biol. Chem.
277, 44202-44207
|Abstract »|Full Text »|PDF »
Broad Requirement for the Mediator Subunit RGR-1 for Transcription in the Caenorhabditis elegans Embryo.
E. Y. Shim, A. K. Walker, and T. K. Blackwell (2002)
J. Biol. Chem.
277, 30413-30416
|Abstract »|Full Text »|PDF »
Novel Mediator Proteins of the Small Mediator Complex in Drosophila SL2 Cells.
J.-Y. Gu, J. M. Park, E. J. Song, G. Mizuguchi, J. H. Yoon, J. Kim-Ha, K.-J. Lee, and Y.-J. Kim (2002)
J. Biol. Chem.
277, 27154-27161
|Abstract »|Full Text »|PDF »
TFIID and human mediator coactivator complexes assemble cooperatively on promoter DNA.
K. M. Johnson, J. Wang, A. Smallwood, C. Arayata, and M. Carey (2002)
Genes & Dev.
16, 1852-1863
|Abstract »|Full Text »|PDF »
Human CRSP interacts with RNA polymerase II CTD and adopts a specific CTD-bound conformation.
A. M. Naar, D. J. Taatjes, W. Zhai, E. Nogales, and R. Tjian (2002)
Genes & Dev.
16, 1339-1344
|Abstract »|Full Text »|PDF »
