Repression of basal transcription by HMG2 is counteracted by TFIIH-associated factors in an ATP-dependent process

A basal repressor of class II gene transcription was identified, purified, and found to be identical to nonhistone chromosomal protein HMG2. HMG2 was shown to inhibit basal transcription under conditions in which transcription templates form soluble complexes with HMG2. Order-of-addition experiments clearly revealed that HMG2 acted after assembly of a TBP-TFIIA-promoter complex and before formation of the fourth phosphodiester bond by RNA polymerase II. Subsequently, an activity that efficiently counteracted repression of transcription by HMG2 in both TBP- and TFIID-containing transcription systems was isolated. Several lines of evidence suggested that antirepression was mediated by a TFIIH-associated factor. The antirepressor first coeluted with TFIIH, was depleted from this fraction by antibodies directed against the TFIIH subunit p62, was dependent on either ATP or dATP, and then was inhibited by the ATP analogs AMP-PNP and ATP gamma S. Relief of HMG2-mediated repression as well as basal promoter function of TFIIH may involve a helicase that coelutes with TFIIH and displays similar nucleotide specificities. Taken together, these data suggest novel consequences of chromatin-associated HMG proteins and they provide direct evidence for a role of TFIIH-associated enzymes in ATP-dependent antirepression of nonhistone chromosomal proteins.

[1]  R. Roeder,et al.  RNA polymerase II cofactor PC2 facilitates activation of transcription by GAL4-AH in vitro , 1994, Molecular and cellular biology.

[2]  D. Reinberg,et al.  DNA topoisomerase I is involved in both repression and activation of transcription , 1993, Nature.

[3]  R. C. Johnson,et al.  The nonspecific DNA-binding and -bending proteins HMG1 and HMG2 promote the assembly of complex nucleoprotein structures. , 1993, Genes & development.

[4]  J. T. Kadonaga,et al.  Identification of a minimal set of proteins that is sufficient for accurate initiation of transcription by RNA polymerase II. , 1993, Genes & development.

[5]  A. Hoffmann,et al.  Unique TATA‐binding protein‐containing complexes and cofactors involved in transcription by RNA polymerases II and III. , 1993, The EMBO journal.

[6]  D. Reinberg,et al.  Multiple functional domains of human transcription factor IIB: distinct interactions with two general transcription factors and RNA polymerase II. , 1993, Genes & development.

[7]  R. Conaway,et al.  Phosphorylation of C-terminal domain of RNA polymerase II is not required in basal transcription , 1993, Nature.

[8]  P. Sharp,et al.  DNA topology and a minimal set of basal factors for transcription by RNA polymerase II , 1993, Cell.

[9]  D. Auble,et al.  An ATP-dependent inhibitor of TBP binding to DNA. , 1993, Genes & development.

[10]  P. Chambon,et al.  DNA repair helicase: a component of BTF2 (TFIIH) basic transcription factor. , 1993, Science.

[11]  T. Boyer,et al.  Factors (TAFs) required for activated transcription interact with TATA box-binding protein conserved core domain. , 1993, Genes & development.

[12]  R. Conaway,et al.  Multifunctional RNA Polymerase I1 Initiation Factor 6 from Rat Liver , 1993 .

[13]  D. Reinberg,et al.  Initiation of transcription by RNA polymerase II: a multi-step process. , 1993, Progress in nucleic acid research and molecular biology.

[14]  R. Conaway,et al.  General initiation factors for RNA polymerase II. , 1993, Annual review of biochemistry.

[15]  M. Horikoshi,et al.  TFIIA induces conformational changes in TFIID via interactions with the basic repeat , 1992, Molecular and cellular biology.

[16]  Qiang Zhou,et al.  Holo-TFIID supports transcriptional stimulation by diverse activators and from a TATA-less promoter. , 1992, Genes & development.

[17]  P. Chambon,et al.  Cloning of the 62-kilodalton component of basic transcription factor BTF2. , 1992, Science.

[18]  J. Gralla,et al.  The acidic activator GAL4-AH can stimulate polymerase II transcription by promoting assembly of a closed complex requiring TFIID and TFIIA. , 1992, Genes & development.

[19]  D. Reinberg,et al.  Factors involved in specific transcription by mammalian RNA polymerase II: purification and analysis of transcription factor IIA and identification of transcription factor IIJ , 1992, Molecular and cellular biology.

[20]  R. Roeder,et al.  Family of proteins that interact with TFIID and regulate promoter activity , 1991, Cell.

[21]  R. Tjian,et al.  Isolation of coactivators associated with the TATA-binding protein that mediate transcriptional activation , 1991, Cell.

[22]  K. Khrapko,et al.  Distribution of high mobility group proteins 1/2, E and 14/17 and linker histones H1 and H5 on transcribed and non-transcribed regions of chicken erythrocyte chromatin. , 1991, Nucleic acids research.

[23]  M. Horikoshi,et al.  Factors involved in specific transcription by mammalian RNA polymerase II: identification of general transcription factor TFIIG. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[24]  D. Reinberg,et al.  Factors involved in specific transcription by mammalian RNA polymerase II: role of transcription factors IIA, IID, and IIB during formation of a transcription-competent complex , 1990, Molecular and cellular biology.

[25]  A. V. D. Eb,et al.  A presumed DNA helicase encoded by ERCC-3 is involved in the human repair disorders xeroderma pigmentosum and Cockayne's syndrome , 1990, Cell.

[26]  D. Landsman,et al.  Structural features of the HMG chromosomal proteins and their genes. , 1990, Biochimica et biophysica acta.

[27]  D. Reinberg,et al.  Factors involved in specific transcription by mammalian RNA polymerase II. Purification and subunit composition of transcription factor IIF. , 1990, The Journal of biological chemistry.

[28]  F. Studier,et al.  Use of T7 RNA polymerase to direct expression of cloned genes. , 1990, Methods in enzymology.

[29]  R. Conaway,et al.  An RNA polymerase II transcription factor has an associated DNA-dependent ATPase (dATPase) activity strongly stimulated by the TATA region of promoters. , 1989, Proceedings of the National Academy of Sciences of the United States of America.

[30]  P. Sharp,et al.  Five intermediate complexes in transcription initiation by RNA polymerase II , 1989, Cell.

[31]  W. Kim,et al.  Purification of RNA polymerase IIO from calf thymus. , 1988, The Journal of biological chemistry.

[32]  M. Horikoshi,et al.  Factors involved in specific transcription by mammalian RNA polymerase II: purification, genetic specificity, and TATA box-promoter interactions of TFIID , 1988, Molecular and cellular biology.

[33]  F. Watt,et al.  High mobility group proteins 1 and 2 stimulate binding of a specific transcription factor to the adenovirus major late promoter. , 1988, Nucleic acids research.

[34]  R. Roeder,et al.  Interaction of a gene-specific transcription factor with the adenovirus major late promoter upstream of the TATA box region , 1985, Cell.

[35]  A. Klug,et al.  Structure of the nucleosome core particle at 7 Å resolution , 1984, Nature.

[36]  L. Kuehl,et al.  Concentrations of high-mobility-group proteins in the nucleus and cytoplasm of several rat tissues , 1984, The Journal of cell biology.

[37]  R. Rill,et al.  Enrichment of transcribed and newly replicated DNA in soluble chromatin released from nuclei by mild micrococcal nuclease digestion. , 1984, Biochimica et biophysica acta.

[38]  J. Palau,et al.  Interaction between domains in chromosomal protein HMG‐1. , 1984, The EMBO journal.

[39]  J. Palau,et al.  DNA and histone H1 interact with different domains of HMG 1 and 2 proteins. , 1983, The EMBO journal.

[40]  E. W. Johns The HMG chromosomal proteins , 1982 .

[41]  F. Eckstein,et al.  Nucleoside phosphorothioates. , 1970, Journal of the American Chemical Society.