Molecular architecture of the S. cerevisiae SAGA complex.

The Saccharomyces cerevisiae SAGA complex is a multifunctional coactivator that regulates transcription by RNA polymerase II. The 3D structure of SAGA, revealed by electron microscopy, is formed by five modular domains and shows a high degree of structural conservation to human TFTC, reflecting their related subunit composition. The positions of several SAGA subunits were mapped by immunolabeling and by analysis of mutant complexes. The Taf (TBP-associated factor) subunits, shared with TFIID, occupy a central region in SAGA and form a similar structure in both complexes. The locations of two histone fold-containing core subunits, Spt7 and Ada1, are consistent with their role in providing a SAGA-specific interface with the Tafs. Three components that perform distinct regulatory functions, Spt3, Gcn5, and Tra1, are spatially separated, underscoring the modular nature of the complex. Our data provide insights into the molecular architecture of SAGA and imply a functional organization to the complex.

[1]  S. Sanders,et al.  TAF25p, a Non-histone-like Subunit of TFIID and SAGA Complexes, Is Essential for Total mRNA Gene Transcription in Vivo * , 1999, The Journal of Biological Chemistry.

[2]  D. Stillman,et al.  Regulation of TATA-Binding Protein Binding by the SAGA Complex and the Nhp6 High-Mobility Group Protein , 2003, Molecular and Cellular Biology.

[3]  J. Workman,et al.  Recruitment of HAT Complexes by Direct Activator Interactions with the ATM-Related Tra1 Subunit , 2001, Science.

[4]  M van Heel,et al.  A new generation of the IMAGIC image processing system. , 1996, Journal of structural biology.

[5]  Fred Winston,et al.  Construction of a set of convenient saccharomyces cerevisiae strains that are isogenic to S288C , 1995, Yeast.

[6]  Fred Winston,et al.  Methods in Yeast Genetics: A Laboratory Course Manual , 1990 .

[7]  B. Chait,et al.  Human STAGA Complex Is a Chromatin-Acetylating Transcription Coactivator That Interacts with Pre-mRNA Splicing and DNA Damage-Binding Factors In Vivo , 2001, Molecular and Cellular Biology.

[8]  F. Winston,et al.  Analysis of Spt7 Function in the Saccharomyces cerevisiae SAGA Coactivator Complex , 2002, Molecular and Cellular Biology.

[9]  Fred Winston,et al.  Functional Organization of the Yeast SAGA Complex: Distinct Components Involved in Structural Integrity, Nucleosome Acetylation, and TATA-Binding Protein Interaction , 1999, Molecular and Cellular Biology.

[10]  L. Guarente,et al.  ADA1, a novel component of the ADA/GCN5 complex, has broader effects than GCN5, ADA2, or ADA3 , 1997, Molecular and cellular biology.

[11]  Jun Qin,et al.  Histone-like TAFs within the PCAF Histone Acetylase Complex , 1998, Cell.

[12]  D. Moras,et al.  The Human TFIID Components TAFII135 and TAFII20 and the Yeast SAGA Components ADA1 and TAFII68 Heterodimerize to Form Histone-Like Pairs , 2000, Molecular and Cellular Biology.

[13]  D. Sterner,et al.  The SANT Domain of Ada2 Is Required for Normal Acetylation of Histones by the Yeast SAGA Complex* , 2002, The Journal of Biological Chemistry.

[14]  Michael R. Green,et al.  Redundant roles for the TFIID and SAGA complexes in global transcription , 2000, Nature.

[15]  Jerry L. Workman,et al.  Expanded Lysine Acetylation Specificity of Gcn5 in Native Complexes* , 1999, The Journal of Biological Chemistry.

[16]  R. Tjian,et al.  Human CRSP interacts with RNA polymerase II CTD and adopts a specific CTD-bound conformation. , 2002, Genes & development.

[17]  J. Strathern,et al.  Methods in yeast genetics : a Cold Spring Harbor Laboratory course manual , 2005 .

[18]  Marin van Heel,et al.  Similarity measures between images , 1987 .

[19]  Andrew J. Link,et al.  Proteomics of the Eukaryotic Transcription Machinery: Identification of Proteins Associated with Components of Yeast TFIID by Multidimensional Mass Spectrometry , 2002, Molecular and Cellular Biology.

[20]  R Ohba,et al.  Yeast Gcn5 functions in two multisubunit complexes to acetylate nucleosomal histones: characterization of an Ada complex and the SAGA (Spt/Ada) complex. , 1997, Genes & development.

[21]  P. Grant,et al.  NuA4, an essential transcription adaptor/histone H4 acetyltransferase complex containing Esa1p and the ATM‐related cofactor Tra1p , 1999, The EMBO journal.

[22]  M. Heel,et al.  Angular reconstitution: a posteriori assignment of projection directions for 3D reconstruction. , 1987 .

[23]  C. Allis,et al.  Histone acetyltransferases. , 2001, Annual review of biochemistry.

[24]  L. Tora,et al.  Histone Folds Mediate Selective Heterodimerization of Yeast TAFII25 with TFIID Components yTAFII47 and yTAFII65 and with SAGA Component ySPT7 , 2001, Molecular and Cellular Biology.

[25]  F. Winston,et al.  Essential functional interactions of SAGA, a Saccharomyces cerevisiae complex of Spt, Ada, and Gcn5 proteins, with the Snf/Swi and Srb/mediator complexes. , 1997, Genetics.

[26]  Michael R Green,et al.  In vivo target of a transcriptional activator revealed by fluorescence resonance energy transfer. , 2004, Genes & development.

[27]  M. Cosma.  Ordered recruitment: gene-specific mechanism of transcription activation. , 2002, Molecular cell.

[28]  D. Sterner,et al.  Inhibition of TATA-Binding Protein Function by SAGA Subunits Spt3 and Spt8 at Gcn4-Activated Promoters , 2000, Molecular and Cellular Biology.

[29]  H. Reinke,et al.  Multiple Mechanistically Distinct Functions of SAGA at the PHO5 Promoter , 2003, Molecular and Cellular Biology.

[30]  F. Winston,et al.  The S. cerevisiae SAGA complex functions in vivo as a coactivator for transcriptional activation by Gal4. , 2001, Genes & development.

[31]  Zhengjian Zhang,et al.  Identification of a Yeast Transcription Factor IID Subunit, TSG2/TAF48* , 2000, The Journal of Biological Chemistry.

[32]  M. Green,et al.  SAGA is an essential in vivo target of the yeast acidic activator Gal4p. , 2001, Genes & development.

[33]  P Hiselius,et al.  The New Generation , 2021, The Women's Liberation Movement in Russia.

[34]  S. Sanders,et al.  Use of a Genetically Introduced Cross-linker to Identify Interaction Sites of Acidic Activators within Native Transcription Factor IID and SAGA* , 2003, The Journal of Biological Chemistry.

[35]  K. Yamamoto,et al.  Identification of TATA-binding Protein-free TAFII-containing Complex Subunits Suggests a Role in Nucleosome Acetylation and Signal Transduction* , 1999, The Journal of Biological Chemistry.

[36]  M. Brand,et al.  Three-dimensional structures of the TAFII-containing complexes TFIID and TFTC. , 1999, Science.

[37]  Robert Pepperell,et al.  What is a Human? , 1994, Intell. Tutoring Media.

[38]  A. Hoffmann,et al.  A histone octamer-like structure within TFIID , 1996, Nature.

[39]  C. Allis,et al.  Tetrahymena Histone Acetyltransferase A: A Homolog to Yeast Gcn5p Linking Histone Acetylation to Gene Activation , 1996, Cell.

[40]  J. Reese,et al.  Transcription Allele-Specific and Broad Defects in Mutants Displaying TAF 90 Analysis of , 2001 .

[41]  Steven L. Cohen,et al.  Structural similarity between TAFs and the heterotetrameric core of the histone octamer , 1996, Nature.

[42]  L. Tora,et al.  Mapping histone fold TAFs within yeast TFIID , 2002, The EMBO journal.

[43]  G. Fink,et al.  Laboratory course manual for methods in yeast genetics , 1986 .

[44]  John R Yates,et al.  A Subset of TAFIIs Are Integral Components of the SAGA Complex Required for Nucleosome Acetylation and Transcriptional Stimulation , 1998, Cell.

[45]  L. Tora,et al.  Distinct Mutations in Yeast TAFII25 Differentially Affect the Composition of TFIID and SAGA Complexes as Well as Global Gene Expression Patterns , 2002, Molecular and Cellular Biology.

[46]  K. Struhl,et al.  The VP16 Activation Domain Interacts with Multiple Transcriptional Components as Determined by Protein-Protein Cross-linking in Vivo * , 2002, The Journal of Biological Chemistry.

[47]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[48]  J R Yates,et al.  The ATM-related cofactor Tra1 is a component of the purified SAGA complex. , 1998, Molecular cell.

[49]  P. Grant,et al.  Role of the Ada2 and Ada3 Transcriptional Coactivators in Histone Acetylation* , 2002, The Journal of Biological Chemistry.

[50]  Robert Tjian,et al.  Structure, Function, and Activator-Induced Conformations of the CRSP Coactivator , 2002, Science.

[51]  R. Roeder,et al.  A Human SPT3-TAFII31-GCN5-L Acetylase Complex Distinct from Transcription Factor IID* , 1998, The Journal of Biological Chemistry.

[52]  L. Tora,et al.  Mapping key functional sites within yeast TFIID , 2004, The EMBO journal.

[53]  J. Reese,et al.  Analysis of TAF90 Mutants Displaying Allele-Specific and Broad Defects in Transcription , 2001, Molecular and Cellular Biology.

[54]  John R. Yates,et al.  Tra1p Is a Component of the Yeast Ada·Spt Transcriptional Regulatory Complexes* , 1998, The Journal of Biological Chemistry.

[55]  J. W. Rooney,et al.  SPT3 interacts with TFIID to allow normal transcription in Saccharomyces cerevisiae. , 1992, Genes & development.

[56]  F. Winston,et al.  SPT20/ADA5 encodes a novel protein functionally related to the TATA-binding protein and important for transcription in Saccharomyces cerevisiae , 1996, Molecular and cellular biology.

[57]  Song Tan,et al.  A histone fold TAF octamer within the yeast TFIID transcriptional coactivator , 2001, Nature Structural Biology.