DNA sequence requirements for generating paused polymerase at the start of hsp 70

Hyunsook Pai, Kevin W. Kraus, Mariana F. Wolfner, John T. Lis

Research output: Contribution to journalArticle

142 Citations (Scopus)

Abstract

RNA polymerase II is transcriptionally engaged but paused &25 nucleotides from the start site of the Asp70 gene of Drosophila melanogaster in uninduced (non-heat-shocked) flies. Hete, we identify regions of the Asp70 promoter that are required for formation of this paused polymerase. Various Asp 70 promoter sequences are substituted for promoter sequences of a yolk protein gene, yp1, which, in males, is normally not expressed and has no paused polymerase. Run-on assays with nuclei of male transgenic flies are used to measure the level of paused polymerase on the hybrid genes. Sequences that reside upstream of the Asp 70 TATA element, when fused upstream of the yp1 TATA element, specify the formation of a paused polymerase on the 5′ end of this hybrid gene. Within this region are multiple copies of the GAGA element, which is known to bind a constitutively expressed factor. This element appears to play a role in generating the pause. Also, in the absence of much of this upstream region, Asp 70 sequences in the vicinity of the transcriptional start and pause site participate in specifying the pause. Deletions of the pause site reduce the level of paused polymerase but do not lead to constitutive transcription. However, a connection between transcription and pausing is seen. The level of paused polymerase on the various hybrid hsp70-yp1 promoters correlates with the promoter's potential to direct heat-induced transcription.

Original languageEnglish
Pages (from-to)284-295
Number of pages12
JournalGenes and Development
Volume6
Issue number2
DOIs
Publication statusPublished - 1992 Jan 1

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Diptera
Genes
Egg Proteins
RNA Polymerase II
Drosophila melanogaster
Genetic Promoter Regions
Nucleotides
Hot Temperature

All Science Journal Classification (ASJC) codes

  • Genetics
  • Developmental Biology

Cite this

Pai, Hyunsook ; Kraus, Kevin W. ; Wolfner, Mariana F. ; Lis, John T. / DNA sequence requirements for generating paused polymerase at the start of hsp 70. In: Genes and Development. 1992 ; Vol. 6, No. 2. pp. 284-295.
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DNA sequence requirements for generating paused polymerase at the start of hsp 70. / Pai, Hyunsook; Kraus, Kevin W.; Wolfner, Mariana F.; Lis, John T.

In: Genes and Development, Vol. 6, No. 2, 01.01.1992, p. 284-295.

Research output: Contribution to journalArticle

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AB - RNA polymerase II is transcriptionally engaged but paused &25 nucleotides from the start site of the Asp70 gene of Drosophila melanogaster in uninduced (non-heat-shocked) flies. Hete, we identify regions of the Asp70 promoter that are required for formation of this paused polymerase. Various Asp 70 promoter sequences are substituted for promoter sequences of a yolk protein gene, yp1, which, in males, is normally not expressed and has no paused polymerase. Run-on assays with nuclei of male transgenic flies are used to measure the level of paused polymerase on the hybrid genes. Sequences that reside upstream of the Asp 70 TATA element, when fused upstream of the yp1 TATA element, specify the formation of a paused polymerase on the 5′ end of this hybrid gene. Within this region are multiple copies of the GAGA element, which is known to bind a constitutively expressed factor. This element appears to play a role in generating the pause. Also, in the absence of much of this upstream region, Asp 70 sequences in the vicinity of the transcriptional start and pause site participate in specifying the pause. Deletions of the pause site reduce the level of paused polymerase but do not lead to constitutive transcription. However, a connection between transcription and pausing is seen. The level of paused polymerase on the various hybrid hsp70-yp1 promoters correlates with the promoter's potential to direct heat-induced transcription.

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