SUMO-1 Modification of Human Transcription Factor (TF) IID Complex Subunits

Abstract

The TFIID complex is composed of the TATA-binding protein (TBP) and TBP-associated factors (TAFs) and is the only component of the general RNA polymerase II (RNAP II) transcription machinery with intrinsic sequence-specific DNA-binding activity. Binding of transcription factor (TF) IID to the core promoter region of protein-coding genes is a key event in RNAP II transcription activation and is the first and rate-limiting step of transcription initiation complex assembly. Intense research efforts in the past have established that TFIID promoter-binding activity as well as the function of TFIID-promoter complexes is tightly regulated through dynamic TFIID interactions with positive- and negative-acting transcription regulatory proteins. However, very little is known about the role of post-translational modifications in the regulation of TFIID. Here we show that the human TFIID subunits hsTAF5 and hsTAF12 are modified by the small ubiquitin-related modifier SUMO-1 in vitro and in human cells. We identify Lys-14 in hsTAF5 and Lys-19 in hsTAF12 as the primary SUMO-1 acceptor sites and show that SUMO conjugation has no detectable effect on nuclear import or intranuclear distribution of hsTAF5 and hsTAF12. Finally, we demonstrate that purified human TFIID complex can be SUMO-1-modified in vitro at both hsTAF5 and hsTAF12. We find that SUMO-1 conjugation at hsTAF5 interferes with binding of TFIID to promoter DNA, whereas modification of hsTAF12 has no detectable effect on TFIID promoter-binding activity. Our observations suggest that reversible SUMO modification at hsTAF5 contributes to the dynamic regulation of TFIID promoter-binding activity in human cells. The TFIID complex is composed of the TATA-binding protein (TBP) and TBP-associated factors (TAFs) and is the only component of the general RNA polymerase II (RNAP II) transcription machinery with intrinsic sequence-specific DNA-binding activity. Binding of transcription factor (TF) IID to the core promoter region of protein-coding genes is a key event in RNAP II transcription activation and is the first and rate-limiting step of transcription initiation complex assembly. Intense research efforts in the past have established that TFIID promoter-binding activity as well as the function of TFIID-promoter complexes is tightly regulated through dynamic TFIID interactions with positive- and negative-acting transcription regulatory proteins. However, very little is known about the role of post-translational modifications in the regulation of TFIID. Here we show that the human TFIID subunits hsTAF5 and hsTAF12 are modified by the small ubiquitin-related modifier SUMO-1 in vitro and in human cells. We identify Lys-14 in hsTAF5 and Lys-19 in hsTAF12 as the primary SUMO-1 acceptor sites and show that SUMO conjugation has no detectable effect on nuclear import or intranuclear distribution of hsTAF5 and hsTAF12. Finally, we demonstrate that purified human TFIID complex can be SUMO-1-modified in vitro at both hsTAF5 and hsTAF12. We find that SUMO-1 conjugation at hsTAF5 interferes with binding of TFIID to promoter DNA, whereas modification of hsTAF12 has no detectable effect on TFIID promoter-binding activity. Our observations suggest that reversible SUMO modification at hsTAF5 contributes to the dynamic regulation of TFIID promoter-binding activity in human cells. To initiate mRNA synthesis, RNA polymerase II (RNAP II) 1The abbreviations used are: RNAP II, RNA polymerase II; HSV-1, herpes simplex virus 1; PIC, preinitiation complex; TBP, TATA-binding protein; TAF, TBP-associated factor; TF, transcription factor; E1, ubiquitin-activating enzyme; E2, ubiquitin carrier protein; E3, ubiquitin-protein isopeptide ligase; HA, hemagglutinin; (HA)3, triple-HA epitope tag; aa, amino acid(s); NEM, N-ethylmaleimide; Ni-NTA, nickel-nitrilotriacetic acid; TBS, Tris-buffered saline; PBS, phosphate-buffered saline; f:TFIID, FLAG:epitope-tagged TFIID complex; ATPγS, adenosine 5′-O-(thiotriphosphate).1The abbreviations used are: RNAP II, RNA polymerase II; HSV-1, herpes simplex virus 1; PIC, preinitiation complex; TBP, TATA-binding protein; TAF, TBP-associated factor; TF, transcription factor; E1, ubiquitin-activating enzyme; E2, ubiquitin carrier protein; E3, ubiquitin-protein isopeptide ligase; HA, hemagglutinin; (HA)3, triple-HA epitope tag; aa, amino acid(s); NEM, N-ethylmaleimide; Ni-NTA, nickel-nitrilotriacetic acid; TBS, Tris-buffered saline; PBS, phosphate-buffered saline; f:TFIID, FLAG:epitope-tagged TFIID complex; ATPγS, adenosine 5′-O-(thiotriphosphate). must assemble in an ordered fashion with a set of general transcription factors, TFIIA, -IIB, -IID, -IIE, -IIF, and -IIH, to form a so-called preinitiation complex (PIC) at the core promoter region of protein-coding genes (1Roeder R.G. Trends Biochem. Sci. 1996; 21: 327-335Abstract Full Text PDF PubMed Scopus (718) Google Scholar, 2Lee T.I. Young R.A. Annu. Rev. Genet. 2000; 34: 77-137Crossref PubMed Scopus (625) Google Scholar, 3Orphanides G. Lagrange T. Reinberg D. Genes Dev. 1996; 10: 2657-2683Crossref PubMed Scopus (844) Google Scholar). The initial, and rate-limiting, step for PIC assembly is binding of TFIID, a multiprotein complex composed of the TATA-binding protein TBP and at least 13 TBP-associated factors (TAFs), to core promoter sequence elements (1Roeder R.G. Trends Biochem. Sci. 1996; 21: 327-335Abstract Full Text PDF PubMed Scopus (718) Google Scholar, 2Lee T.I. Young R.A. Annu. Rev. Genet. 2000; 34: 77-137Crossref PubMed Scopus (625) Google Scholar, 3Orphanides G. Lagrange T. Reinberg D. Genes Dev. 1996; 10: 2657-2683Crossref PubMed Scopus (844) Google Scholar, 4Gangloff Y. Romier C. Thuault S. Werten S. Davidson I. Trends Biochem. Sci. 2001; 26: 250-257Abstract Full Text Full Text PDF PubMed Scopus (114) Google Scholar, 5Tora L. Genes Dev. 2002; 16: 673-675Crossref PubMed Scopus (192) Google Scholar). In addition to sequence-specific DNA binding, the TFIID complex provides several enzymatic activities that all reside in its largest subunit TAF1 (6Wassarman D.A. Sauer F. J. Cell Sci. 2001; 114: 2895-2902Crossref PubMed Google Scholar). These include histone acetyltransferase activity (7Mizzen C.A. Yang X.J. Kokubo T. Brownell J.E. Bannister A.J. Owen-Hughes T. Workman J. Wang L. Berger S.L. Kouzarides T. Nakatani Y. Allis C.D. Cell. 1996; 87: 1261-1270Abstract Full Text Full Text PDF PubMed Scopus (617) Google Scholar), protein kinase activity (8Dikstein R. Ruppert S. Tjian R. Cell. 1996; 84: 781-790Abstract Full Text Full Text PDF PubMed Scopus (170) Google Scholar), and ubiquitin-activating/-conjugating activity (9Pham A.-D. Sauer F. Science. 2000; 289: 2357-2360Crossref PubMed Scopus (203) Google Scholar). How exactly TFIID enzymatic activities contribute to RNAP II transcription is still unknown. In accordance with its crucial role in PIC assembly, TFIID is considered one of the key targets of transcription regulation pathways. Biochemical and genetic studies have shown that TFIID binding to promoters as well as the stability and functionality of TFIID-promoter complexes are subject to regulation by gene-specific activators and repressors (10Hoffmann A. Oelgeschläger T. Roeder R.G. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 8928-8935Crossref PubMed Scopus (76) Google Scholar, 11Roeder R.G. Cold Spring Harbor Symp. Quant. Biol. 1998; 63: 201-218Crossref PubMed Scopus (141) Google Scholar) and by an array of ubiquitous regulators of TFIID (TBP) activity, including NC2 (Dr1/DRAP1), Mot1/BTAF1, and the NOT complex (11Roeder R.G. Cold Spring Harbor Symp. Quant. Biol. 1998; 63: 201-218Crossref PubMed Scopus (141) Google Scholar, 12Pugh B.F. Gene (Amst.). 2000; 255: 1-14Crossref PubMed Scopus (154) Google Scholar, 13Lee T.I. Young R.A. Genes Dev. 1998; 12: 1398-1408Crossref PubMed Scopus (155) Google Scholar, 14Pereira L.A. Klejman M.P. Timmers H.T. Gene (Amst.). 2003; 315: 1-13Crossref PubMed Scopus (43) Google Scholar). In addition, TFIID subunits are subject to post-transcriptional modifications. Earliest studies demonstrated that several subunits of TFIID are phosphorylated during mitosis and that TFIID isolated from mitotic cells fails to respond to transcription activators in vitro (15Segil N. Guermah M. Hoffmann A. Roeder R.G. Heintz N. Genes Dev. 1996; 10: 2389-2400Crossref PubMed Scopus (160) Google Scholar). Results of more recent studies demonstrated gene-specific effects upon hsTAF10 methylation by the SET9 protein methyltransferase (16Kouskouti A. Scheer E. Staub A. Tora L. Talianidis I. Mol. Cell. 2004; 14: 175-182Abstract Full Text Full Text PDF PubMed Scopus (207) Google Scholar) and that the C-terminal domain of TAF1 is a substrate for the protein kinase CKII (17Sawa C. Nedea E. Krogan N. Wada T. Handa H. Greenblatt J. Buratowski S. Mol. Cell. Biol. 2004; 24: 4734-4742Crossref PubMed Scopus (30) Google Scholar). Exactly how and to what extent these TAF modifications modulate TFIID functions is not known. In recent years, regulation of transcription factor activity through modification with SUMO proteins has attracted considerable attention (18Seeler J.S. Dejean A. Nat. Rev. Mol. Cell. Biol. 2003; 4: 690-699Crossref PubMed Scopus (575) Google Scholar, 19Verger A. Perdomo J. Crossley M. EMBO Rep. 2003; 4: 137-142Crossref PubMed Scopus (369) Google Scholar, 20Gill G. Genes Dev. 2004; 18: 2046-2059Crossref PubMed Scopus (618) Google Scholar, 21Gill G. Curr. Opin. Genet. Dev. 2003; 13: 108-113Crossref PubMed Scopus (193) Google Scholar). SUMO proteins are small ubiquitin-related modifiers that are conjugated to target proteins through an enzymatic pathway, which is similar to ubiquitylation, but which involves a distinct set of Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar, F. Annu. Rev. Cell Dev. Biol. 2000; 16: PubMed Scopus Google Scholar). SUMO proteins are as that have to be by to a C-terminal for The form of SUMO is in an in which a is its C-terminal and a in the to a SUMO is to a target protein isopeptide SUMO and are to target proteins in vitro F. Annu. Rev. Cell Dev. Biol. 2000; 16: PubMed Scopus Google Scholar), several SUMO have that of SUMO from the to Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar). SUMO modification is a reversible and dynamic and SUMO can be from substrate proteins by Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar). SUMO modification in but not a is a amino and is amino Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar, C. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). Here we SUMO-1 modification of several TFIID subunits the We show that hsTAF5 and hsTAF12 are SUMO-1-modified in human cells. We show that and can be SUMO-1-modified a SUMO conjugation composed of and and and can be in purified TFIID whereas SUMO acceptor sites in TAF1 and TBP to upon assembly TFIID. We the SUMO-1 acceptor sites in hsTAF5 and hsTAF12 and show that SUMO modification not nuclear import or the nuclear distribution of hsTAF5 and hsTAF12 in human cells. Finally, we of in vitro DNA binding that SUMO modification of purified human TFIID at hsTAF5 TFIID DNA-binding activity. Cell cells in modified with SUMO-1 cells D. J. 2002; PubMed Scopus Google Scholar) by and in the of to for for and a of R. G. Roeder the of has G. Cell. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar). for the and as well as for the from binding sites for the in of a core promoter with a of the transcription and by the from E. Oelgeschläger T. A.J. Roeder R.G. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar) at and and used for protein by in vitro and by the for a triple-HA epitope amino sequence to the of or hsTAF12 A. Roeder R.G. J. Biol. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, Y. Guermah M. E. Oelgeschläger T. S. R. T. M. Roeder R.G. J. Biol. 1997; Full Text Full Text PDF PubMed Scopus Google Scholar, S. R. Nakatani Y. M. Roeder R.G. PubMed Scopus Google Scholar), at and and and used to hsTAF5 and hsTAF12 in cells. for and by and and used to in human cells of the form of to the of hsTAF5 the first 13 amino or hsTAF12 the first amino To of the SUMO-1 from the proteins by in the C-terminal of the form of SUMO-1 to used for of form of SUMO-1 in To the for from D. J. 2002; PubMed Scopus Google Scholar) and and for TBP and and from G. Roeder of and with or cells in at and the with in modified at a of of The virus to to cells for at with and the cells at in and by and Cold of SUMO-1 cells in at The cells to for at for cells in and by and to the in with cells and of of from Cell by at in of with and for at and for at with in of to proteins conjugated to or with in of to FLAG:epitope-tagged proteins. binding, the with of NEM, in the of and with of with or of the by and and by and to and an in for with to the with in and with primary in for in and for with the in for in and with cells on and in for for in PBS, with in for for in PBS, and with in for for with primary in for with PBS, and for with in with and on a an and and of by in vitro for at in of the of and or of and the and of hsTAF12 and purified as A. Roeder R.G. J. Biol. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar, T. Y. Roeder R.G. Mol. Cell. 1998; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar, E. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). The form of human SUMO-1 in E. and purified from on to the FLAG:epitope-tagged TFIID complex purified by from the TFIID as H. Wang Hoffmann A. Roeder R.G. EMBO J. 12: PubMed Scopus Google Scholar). In SUMO-1 conjugation in an or in of purified of purified of purified human and of purified protein substrate or of in vitro at or for as of DNA promoter DNA by from a purified from and on to the TFIID DNA Binding DNA binding at for in transcription binding of purified TBP or of FLAG:epitope-tagged TFIID and of promoter complexes from proteins in a with of transcription and from with in the and by binding to of for at and with In vitro SUMO-1 conjugation of to or DNA binding in as SUMO-1 hsTAF5 and hsTAF12 by SUMO-1 in sequence that several human TFIID including and the target sequence Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar, C. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar) We to of TFIID subunits in human cells. of SUMO-1 is only a small of SUMO-1 target proteins is modified at Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar). SUMO conjugation is in cells that SUMO as a of SUMO-1 of SUMO targets is by of of SUMO for of SUMO from by To SUMO-1 modification of human TFIID we of a human that of human SUMO-1 protein D. J. 2002; PubMed Scopus Google Scholar). in the of and to and to proteins conjugated to of by with an for of hsTAF5 and hsTAF12 in but not in of cells and SUMO-1-modified but not SUMO-1-modified be in of cells and not we to for SUMO-1 modification of TFIID subunits with to studies that as and by SUMO-1 conjugation in human cells D. J. 2002; PubMed Scopus Google Scholar, Y. R. M. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus (203) Google Scholar, T. M. Y. Y. H. H. S. H. M. Biochem. 2003; PubMed Scopus (43) Google Scholar). To of TFIID subunits in SUMO-1 is we the of hsTAF5 protein in to including and virus at and at no detectable effect on hsTAF5 not In at and with herpes simplex virus in a of SUMO-1 and a of SUMO-1-modified hsTAF5 to a detectable in the of hsTAF5 the of SUMO-1-modified hsTAF5 in to can be to of activity of SUMO activity. In these observations demonstrate that hsTAF5 and hsTAF12 are modified by SUMO-1 in human cells and that TAF SUMO-1 conjugation in SUMO-1 cells is a regulated and hsTAF12 for SUMO-1 in we SUMO-1 modification of TFIID subunits in purified hsTAF12 or or or hsTAF5 by in vitro with a SUMO-1 conjugation composed of purified human and and purified SUMO-1 J. Biol. Full Text Full Text PDF PubMed Scopus Google Scholar, E. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). of SUMO-1 modification of and the of hsTAF12 SUMO-1 modification not with which a SUMO target and with the of hsTAF12 which the of A. Roeder R.G. J. Biol. 1996; Full Text Full Text PDF PubMed Scopus Google Scholar). SUMO-1 target one to a region histone acetyltransferase and ubiquitin-activating/-conjugating activities (7Mizzen C.A. Yang X.J. Kokubo T. Brownell J.E. Bannister A.J. Owen-Hughes T. Workman J. Wang L. Berger S.L. Kouzarides T. Nakatani Y. Allis C.D. Cell. 1996; 87: 1261-1270Abstract Full Text Full Text PDF PubMed Scopus (617) Google Scholar, A.-D. Sauer F. Science. 2000; 289: 2357-2360Crossref PubMed Scopus (203) Google Scholar), and in to the region Tjian R. Science. 2000; PubMed Scopus Google Scholar). To SUMO modification of at a or at we and the protein by in an amino which SUMO target an and ubiquitin-activating/-conjugating and a C-terminal the and C-terminal kinase domain shown in and which SUMO target be whereas which a SUMO target not be These demonstrate that at least SUMO target sites and are with the that at the SUMO target TFIID in SUMO-1-modified at hsTAF5 and but at TAF1 or The shown in demonstrated that and are SUMO in to these TFIID subunits be the TFIID To we in vitro with FLAG:epitope-tagged TFIID complex H. Wang Hoffmann A. Roeder R.G. EMBO J. 12: PubMed Scopus Google Scholar). with for and subunits of hsTAF5 and hsTAF12 In SUMO modification of and not be These demonstrate that and SUMO targets sites are on the of the human TFIID However, SUMO target sites and to be the in vitro of SUMO-1 in hsTAF12 and on hsTAF5 and these the only subunits for which we demonstrate SUMO modification in human cells and in vitro in the of the TFIID To SUMO-1 modification of hsTAF5 and hsTAF12 at the SUMO-1 target sites by amino sequence we in which the SUMO-1 acceptor by and by in vitro to in vitro and by shown in in vitro SUMO modification of and proteins with the proteins. These demonstrate that Lys-14 in hsTAF5 and Lys-19 in hsTAF12 are the primary acceptor sites for SUMO-1 conjugation in To that hsTAF5 Lys-14 and hsTAF12 Lys-19 are sites for SUMO conjugation in human we and hsTAF5 and hsTAF12 proteins in cells D. J. 2002; PubMed Scopus Google Scholar). and at that the not protein stability in human cells not SUMO-1 modification of and proteins with the of in vitro We that SUMO-1 modification of hsTAF5 and hsTAF12 the sequence and that Lys-14 in hsTAF5 and Lys-19 in hsTAF12 are sites of SUMO-1 conjugation in human cells. SUMO-1 hsTAF12 and hsTAF5 studies SUMO modification with the of target proteins to as (18Seeler J.S. Dejean A. Nat. Rev. Mol. Cell. Biol. 2003; 4: 690-699Crossref PubMed Scopus (575) Google Scholar, 19Verger A. Perdomo J. Crossley M. EMBO Rep. 2003; 4: 137-142Crossref PubMed Scopus (369) Google Scholar, 20Gill G. Genes Dev. 2004; 18: 2046-2059Crossref PubMed Scopus (618) Google Scholar, J.S. Dejean A. 2001; PubMed Scopus Google Scholar). To we hsTAF5 and hsTAF12 and and SUMO acceptor in cells and by of cells with that hsTAF5 is in the with a nuclear distribution the of only be in the not However, with in the that nuclear import of hsTAF5 on a we that of the with in the at SUMO acceptor and nuclear similar to that nuclear import of hsTAF5 and hsTAF12 is of SUMO However, only a small of proteins modified and that in the nuclear distribution of proteins and SUMO acceptor not be We to hsTAF5 and hsTAF12 with SUMO-1 to SUMO-1 acceptor the SUMO acceptor in hsTAF5 and hsTAF12 are very to we that SUMO-1 conjugation be by the form of SUMO-1 to the of hsTAF5 the first 13 amino and to hsTAF12 the first amino However, upon in cells these proteins SUMO-1 and or not To by we the C-terminal in the SUMO-1 of the proteins with The and are in and cells not and show nuclear with no to proteins these suggest that SUMO-1 modification not nuclear import or the nuclear distribution of hsTAF5 and hsTAF12. SUMO-1 at DNA-binding of used an promoter DNA to SUMO conjugation the DNA-binding activity of purified TFIID We first human TFIID be SUMO-1-modified to To promoter DNA on a and with purified TFIID-promoter complexes from protein a and with in vitro SUMO-1 conjugation the in vitro and proteins and in by small of TFIID-promoter complexes during the in However, of or the of of the human SUMO conjugation and and not SUMO-1-modified of hsTAF5 and hsTAF12 be in the and These demonstrate that SUMO-1 can be conjugated to hsTAF5 and hsTAF12 in TFIID TFIID-promoter complex stability and not we binding of purified TFIID complex to promoter first to in vitro SUMO-1 by addition of promoter DNA and for to assemble TFIID-promoter TFIID and complexes and in by We that in vitro SUMO conjugation in the of and not To SUMO-1 conjugation during the DNA binding with the promoter DNA and the DNA-binding in the of ATPγS, SUMO-1-modified only be in the TFIID In TFIID and TFIID a similar of SUMO-1-modified and and and the DNA-binding step in the of ATPγS, of SUMO-1-modified be in TFIID-promoter DNA to shown in these can be to SUMO-1 conjugation TFIID-promoter complex However, these the of TFIID SUMO-1-modified to be in the In the of the DNA binding suggest that SUMO-1 modification at TFIID binding to promoter DNA, whereas SUMO-1 modification at has no detectable effect on TFIID DNA-binding activity. In TFIID-promoter complexes can be SUMO-1-modified at both and TFIID complex In we SUMO-1 modification of several human TFIID subunits the SUMO target Annu. Rev. Biochem. 2004; PubMed Scopus Google Scholar, C. J. Biol. 2001; Full Text Full Text PDF PubMed Scopus Google Scholar). We demonstrate that the human TFIID subunits hsTAF5 and hsTAF12 are in human cells and in both in and in the of purified human TFIID. We in vitro of and at least SUMO acceptor one to the and in to the However, we to demonstrate in vitro SUMO conjugation to or to in the of purified human TFIID TFIID at can more for in the of activity, a that has to be in the the of SUMO modification multiprotein complexes as TFIID a In the conjugation of SUMO to a target or modulate TFIID as TAF interactions with regulatory TBP DNA-binding activity or one of the enzymatic activities in the TAF1 However, is that a SUMO modification can the TFIID for by its stability or by its to nuclear the SUMO modification at a target have only very and be to modulate TFIID Finally, we that in TFIID activity be about by SUMO modification of isolated TFIID which in TFIID complex assembly or the of dynamic in TFIID subunit during L. E. L. Davidson I. Mol. Biol. 2001; PubMed Scopus Google Scholar, G. S. L. M. Davidson I. J. Cell Sci. 2003; PubMed Scopus Google Scholar). we have no that SUMO modification the nuclear of TFIID. In human hsTAF5 and hsTAF12 the SUMO acceptor and and proteins a nuclear similar to However, the not to on the in the nuclear distribution of TFIID can be about by SUMO modification of TFIID is in that is the only component of the general RNAP II transcription machinery of sequence-specific DNA TFIID interactions with core promoter DNA elements are crucial for RNAP II transcription initiation complex assembly and are tightly by transcription regulatory pathways. We the effect of on the DNA-binding activity of purified human TFIID complex promoter DNA Our suggest that SUMO-1 conjugation to hsTAF5 interferes with TFIID binding to promoter In SUMO-1 conjugation to hsTAF12 subunit to have no detectable effect on TFIID DNA-binding activity. These observations suggest that reversible SUMO modification at the hsTAF5 subunit can contribute to the dynamic regulation of TFIID promoter-binding activity in human cells. TFIID-promoter complexes can be at both hsTAF5 and hsTAF12 that hsTAF5 interferes with TFIID DNA interactions through a subunit hsTAF5 studies that the in vitro promoter-binding activity of purified human TFIID complex is on sequence-specific TBP interactions with E. Oelgeschläger T. A.J. Roeder R.G. Proc. Natl. Acad. Sci. U. S. A. PubMed Scopus Google Scholar, T. Roeder R.G. 1996; PubMed Scopus Google Scholar, A. Genes Dev. 1997; PubMed Scopus Google Scholar). SUMO modification of TFIID at hsTAF5 for from of purified human and TFIID complexes by and C. S. C. Tora L. EMBO J. 2002; 21: PubMed Scopus Google Scholar, C. S.L. C. Tora L. EMBO J. 2004; PubMed Scopus Google Scholar, F. C. Tjian R. E. Science. PubMed Scopus (114) Google Scholar, M. C. Tora L. Science. PubMed Scopus Google Scholar). These studies a in the of human and TFIID which a of by F. C. Tjian R. E. Science. PubMed Scopus (114) Google Scholar, M. C. Tora L. Science. PubMed Scopus Google Scholar). The of the TFIID complex has by C. S. C. Tora L. EMBO J. 2002; 21: PubMed Scopus Google Scholar, C. S.L. C. Tora L. EMBO J. 2004; PubMed Scopus Google Scholar). TFIID of S.L. Mol. Cell. Biol. 2002; PubMed Scopus Google Scholar), which form the the of the TFIID C. S.L. C. Tora L. EMBO J. 2004; PubMed Scopus Google Scholar). The of the of are both in of whereas the C. Trends Biochem. Sci. 24: Full Text Full Text PDF PubMed Scopus Google Scholar), and C. S. C. Tora L. EMBO J. 2002; 21: PubMed Scopus Google Scholar, C. S.L. C. Tora L. EMBO J. 2004; PubMed Scopus Google Scholar). a complex with through histone domain interactions Y. Romier C. Thuault S. Werten S. Davidson I. Trends Biochem. Sci. 2001; 26: 250-257Abstract Full Text Full Text PDF PubMed Scopus (114) Google Scholar, Werten S. Romier C. L. D. Davidson I. Mol. Cell. Biol. 2000; PubMed Scopus Google Scholar). TFIID complexes can which to and of the TFIID C. S. C. Tora L. EMBO J. 2002; 21: PubMed Scopus Google Scholar). a on the TFIID which the of hsTAF5 and hsTAF12 SUMO target The that SUMO conjugation to the of hsTAF5 in in to TBP, with the that hsTAF5 interferes with TBP DNA of hsTAF12 distinct one in to the and one in a from These observations suggest an of that must be considered the of SUMO modifications at TFIID subunits is SUMO modification of a TFIID subunit distinct the TFIID studies shown that reversible SUMO conjugation the activity of transcription regulatory proteins in human cells (18Seeler J.S. Dejean A. Nat. Rev. Mol. Cell. Biol. 2003; 4: 690-699Crossref PubMed Scopus (575) Google Scholar, 19Verger A. Perdomo J. Crossley M. EMBO Rep. 2003; 4: 137-142Crossref PubMed Scopus (369) Google Scholar, 21Gill G. Curr. Opin. Genet. Dev. 2003; 13: 108-113Crossref PubMed Scopus (193) Google Scholar). Our these observations and the first that SUMO modifications can the activity of the human TFIID a key component of the general RNAP II transcription Our observations are in with of in which several RNAP II subunits as well as subunits of the general transcription factors TFIIA, TFIID, and as SUMO targets U. T. E. J. Biol. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar, J. Biol. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar, H. J. Biol. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar). SUMO modification of general RNAP II transcription factors to be a regulatory that is We and Roeder for We and of the Oelgeschläger for and