YY1 is an initiator sequence-binding protein that directs and activates transcription in vitro

REGULATION of eukaryotic messenger RNA transcription is governed by DNA sequence elements that serve as binding sites for sequence-specific transcription factors1–3. These include upstream and downstream promoter-proximal elements, enhancers, repressers, and silencers, which modulate the rate of specific initiation by RNA polymerase II. In addition, the promoter-proximal region between −45 and +30 (relative to the start of initiation) contains two highly conserved motifs, the TATA sequence at around −30 and CA at +1 (ref. 4). Although the TATA element-binding factor TFIID has been purified and cloned from several organisms and has provided invaluable insight into the process of transcription initiation and its regulation, little is known about factors that interact at the +1 region. We have recently shown that the adeno-associated virus type 2 P5 promoter +1 region (P5 + 1 element) binds transcription factor YY1 (ref. 5). We report here that this sequence is necessary and sufficient for accurate basal transcription. Further, partially purified YY1 can restore basal level transcription from a P5 + 1 element in a HeLa extract depleted for YY1 or a Drosophila embryo extract devoid of YY1 activity, whereas a YYl-specific antibody can block the reactivation. Finally, using electrophoretic mobility shift assay, we have identified YYl-related factors that bind to two other transcription initiators in cellular genes.

[1]  E. Harlow,et al.  Antibodies: A Laboratory Manual , 1988 .

[2]  F. Sanger,et al.  DNA sequencing with chain-terminating inhibitors. , 1977, Proceedings of the National Academy of Sciences of the United States of America.

[3]  D. Baltimore,et al.  Transcriptional activation by Sp1 as directed through TATA or initiator: specific requirement for mammalian transcription factor IID. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[4]  M. Horikoshi,et al.  Repression of HIV-1 transcription by a cellular protein. , 1991, Science.

[5]  D. Goeddel,et al.  Nucleotide sequence of a portion of human chromosome 9 containing a leukocyte interferon gene cluster. , 1982, Journal of molecular biology.

[6]  P J Farnham,et al.  Transcription initiation from the dihydrofolate reductase promoter is positioned by HIP1 binding at the initiation site. , 1990, Molecular and cellular biology.

[7]  L. Maquat,et al.  Minimal sequence and factor requirements for the initiation of transcription from an atypical, TATATAA box-containing housekeeping promoter. , 1990, The Journal of biological chemistry.

[8]  Yang Shi,et al.  Transcriptional repression by YY1, a human GLI-Krüippel-related protein, and relief of repression by adenovirus E1A protein , 1991, Cell.

[9]  Y. Shi,et al.  Adeno-associated virus P5 promoter contains an adenovirus E1A-inducible element and a binding site for the major late transcription factor , 1989, Journal of virology.

[10]  P. Luciw,et al.  Structural arrangements of transcription control domains within the 5'-untranslated leader regions of the HIV-1 and HIV-2 promoters. , 1988, Genes & development.

[11]  R. Roeder,et al.  Accurate transcription initiation by RNA polymerase II in a soluble extract from isolated mammalian nuclei. , 1983, Nucleic acids research.

[12]  M. Ptashne.  How eukaryotic transcriptional activators work , 1988, Nature.

[13]  F. Lee,et al.  Functional analysis of the steroid hormone control region of mouse mammary tumor virus. , 1984, Nucleic acids research.

[14]  M. Botchan,et al.  A promoter with an internal regulatory domain is part of the origin of replication in BPV-1. , 1987, Science.

[15]  S. Banerjee,et al.  Organization of the transcriptional control region of the E1b gene of adenovirus type 5 , 1988, Journal of virology.

[16]  D. Baltimore,et al.  The “initiator” as a transcription control element , 1989, Cell.

[17]  Robert Tjian,et al.  Control of eukaryotic messenger RNA synthesis by sequence-specific DNA-binding proteins , 1985, Nature.

[18]  H. Varmus,et al.  A small region of the mouse mammary tumor virus long terminal repeat confers glucocorticoid hormone regulation on a linked heterologous gene. , 1983, Proceedings of the National Academy of Sciences of the United States of America.

[19]  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.

[20]  H. Diggelmann,et al.  Glucocorticoid regulation of mouse mammary tumor virus: identification of a short essential DNA region. , 1983, The EMBO journal.

[21]  R. Roeder,et al.  Physical analysis of transcription preinitiation complex assembly on a class II gene promoter. , 1988, Science.

[22]  O. Pongs,et al.  In vitro transcription with extracts of nuclei of Drosophila embryos. , 1985, Nucleic acids research.

[23]  G. Hager,et al.  Steroid-dependent interaction of transcription factors with the inducible promoter of mouse mammary tumor virus in vivo , 1987, Cell.

[24]  P Bucher,et al.  Compilation and analysis of eukaryotic POL II promoter sequences. , 1986, Nucleic acids research.

[25]  T. Okamoto,et al.  Transcriptional activation from the long-terminal repeat of human immunodeficiency virus in vitro. , 1990, Virology.

[26]  B. Safer,et al.  Identification and characterization of an adenovirus 2 major late promoter CAP sequence DNA-binding protein. , 1990, The Journal of biological chemistry.

[27]  P. Chambon,et al.  Sequences upstream from the T-A-T-A box are required in vivo and in vitro for efficient transcription from the adenovirus serotype 2 major late promoter. , 1982, Proceedings of the National Academy of Sciences of the United States of America.

[28]  R. Tjian,et al.  Transcriptional regulation in mammalian cells by sequence-specific DNA binding proteins. , 1989, Science.

[29]  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.

[30]  R. Tjian,et al.  Direct interaction between Sp1 and the BPV enhancer E2 protein mediates synergistic activation of transcription , 1991, Cell.

[31]  R. Tjian,et al.  Affinity purification of sequence-specific DNA binding proteins. , 1986, Proceedings of the National Academy of Sciences of the United States of America.