RNA polymerase II interacts with the promoter region of the noninduced hsp70 gene in Drosophila melanogaster cells.

By using a protein-DNA cross-linking method (D. S. Gilmour and J. T. Lis, Mol. Cell. Biol. 5:2009-2018, 1985), we examined the in vivo distribution of RNA polymerase II on the hsp70 heat shock gene in Drosophila melanogaster Schneider line 2 cells. In heat shock-induced cells, a high level of RNA polymerase II was detected on the entire gene, while in noninduced cells, the RNA polymerase II was confined to the 5' end of the hsp70 gene, predominantly between nucleotides -12 and +65 relative to the start of transcription. This association of RNA polymerase II was apparent whether the cross-linking was performed by a 10-min UV irradiation of chilled cells with mercury vapor lamps or by a 40-microsecond irradiation of cells with a high-energy xenon flash lamp. We hypothesize that RNA polymerase II has access to, and a high affinity for, the promoter region of this gene before induction, and this poised RNA polymerase II may be critical in the mechanism of transcription activation.

[1]  Mark Groudine,et al.  A block to elongation is largely responsible for decreased transcription of c-myc in differentiated HL60 cells , 1986, Nature.

[2]  John T. Lis,et al.  Topoisomerase I interacts with transcribed regions in Drosophila cells , 1986, Cell.

[3]  J. Lis,et al.  In vivo interactions of RNA polymerase II with genes of Drosophila melanogaster , 1985, Molecular and cellular biology.

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

[5]  J. Barsoum,et al.  Preferential localization of variant nucleosomes near the 5'-end of the mouse dihydrofolate reductase gene. , 1985, The Journal of biological chemistry.

[6]  J. Gralla,et al.  Interaction of RNA polymerase with lacUV5 promoter DNA during mRNA initiation and elongation. Footprinting, methylation, and rifampicin-sensitivity changes accompanying transcription initiation. , 1985, Journal of molecular biology.

[7]  S. Chen‐Kiang,et al.  Pausing and premature termination of human RNA polymerase II during transcription of adenovirus in vivo and in vitro. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Lis,et al.  High-resolution mapping of DNase I-hypersensitive sites of Drosophila heat shock genes in Drosophila melanogaster and Saccharomyces cerevisiae , 1984, Molecular and cellular biology.

[9]  J. Lengyel,et al.  Transcription, export and turnover of Hsp70 and αβ two Drosophila heat shock genes sharing a 400 nucleotide 5′ upstream region , 1984 .

[10]  J. Lis,et al.  Detecting protein-DNA interactions in vivo: distribution of RNA polymerase on specific bacterial genes. , 1984, Proceedings of the National Academy of Sciences of the United States of America.

[11]  Donald D. Brown The role of stable complexes that repress and activate eucaryotic genes , 1984, Cell.

[12]  Carl Wu Two protein-binding sites in chromatin implicated in the activation of heat-shock genes , 1984, Nature.

[13]  C. S. Parker,et al.  A Drosophila RNA polymerase II transcription factor contains a promoter-region-specific DNA-binding activity , 1984, Cell.

[14]  Robert Entriken,et al.  Escherichia coli promoter sequences predict in vitro RNA polymerase selectivity , 1984, Nucleic Acids Res..

[15]  S. C. Elgin Anatomy of hypersensitive sites. , 1984, Nature.

[16]  R. Weinmann,et al.  In vitro transcription of adenovirus genes. , 1984, Current topics in microbiology and immunology.

[17]  R. Weinmann,et al.  RNA polymerase II ternary transcription complexes generated in vitro. , 1983, Nucleic acids research.

[18]  K. Yamamoto,et al.  DNA sequences bound specifically by glucocorticoid receptor in vitro render a heterologous promoter hormone responsive in vivo , 1983, Cell.

[19]  C. Wu,et al.  A rapid mixing-photocrosslinking technique to study the dynamics of nucleic acid-protein interactions. , 1983, Analytical biochemistry.

[20]  J. M. Velazquez,et al.  Is the major Drosophila heat shock protein present in cells that have not been heat shocked? , 1983, The Journal of cell biology.

[21]  C. Yanofsky Prokaryotic mechanisms in eukaryotes? , 1983, Nature.

[22]  A. Travers Protein contacts for promoter location in eukaryotes. , 1983, Nature.

[23]  S. McKnight,et al.  Transcriptional control signals of a eukaryotic protein-coding gene. , 1982, Science.

[24]  A. Greenleaf,et al.  Immunological studies of RNA polymerase II using antibodies to subunits of Drosophila and wheat germ enzyme. , 1982, The Journal of biological chemistry.

[25]  Sarah C. R. Elgin,et al.  DNAase I-hypersensitive sites of chromatin , 1981, Cell.

[26]  J. Lis,et al.  Cloning and characterization of nine heat-shock-induced mRNAs of Drosophila melanogaster. , 1981, Gene.

[27]  P. Chambon,et al.  Clustering of RNA polymerase B molecules in the 5' moiety of the adult beta-globin gene of hen erythrocytes. , 1981, Nucleic acids research.

[28]  F. Karch,et al.  Extensive regions of homology in front of the two hsp70 heat shock variant genes in Drosophila melanogaster. , 1981, Journal of molecular biology.

[29]  D. Ish-Horowicz,et al.  Evolution of the 87A and 87C heat-shock loci in Drosophila , 1981, Nature.

[30]  E. Craig,et al.  Sequence of three copies of the gene for the major Drosophila heat shock induced protein and their flanking regions , 1980, Cell.

[31]  Carl Wu The 5′ ends of Drosophila heat shock genes in chromatin are hypersensitive to DNase I , 1980, Nature.

[32]  M. Goldschmidt-Clermont,et al.  Two genes for the major heat-shock protein of Drosophila melanogaster arranged as an inverted repeat. , 1980, Nucleic acids research.

[33]  S. Artavanis-Tsakonas,et al.  Physical map of two D. melanogaster DNA segments containing sequences coding for the 70,000 dalton heat shock protein , 1979, Cell.

[34]  E. Southern Detection of specific sequences among DNA fragments separated by gel electrophoresis. , 1975, Journal of molecular biology.

[35]  M J Chamberlin,et al.  The selectivity of transcription. , 1974, Annual review of biochemistry.

[36]  I. Schneider,et al.  Cell lines derived from late embryonic stages of Drosophila melanogaster. , 1972, Journal of embryology and experimental morphology.