Enhancer containing unusual GC box-like sequences on the human choline acetyltransferase gene

Hiroyasu Inoue, E. Edward Baetge, Louis B. Hersh

Research output: Contribution to journalArticlepeer-review

16 Scopus citations

Abstract

The minimum requirement of an enhancer for the human choline acetyltransferase (ChAT) gene was analyzed by mutagenesis and protein-DNA interaction studies. A series of deletion and site-specific mutants were expressed transiently in a rat cholinergic neuroblastoma cell, NS20Y. The results revealed that the distal region between -970 and -941 base pairs (bp) from the transcription start site is essential for efficient transcription of the human ChAT gene. Two GC box-like sequences were located within this region, and they were shown to bind purified Sp1 transcription factor by footprinting and gel retardation analyses. This sequence, however, has an orientational effect and is located approximately 1000 bp from the transcription start site, unusual for the conventional GC box sequence. Furthermore, the sequence between these GC box-like sequences is shown to bind another novel trans-acting factor by gel retardation assay and to be necessary for efficient enhancer activity. On the other hand, gel retardation assays using a nuclear extract from Drosophila SL2 cells suggested that non-Sp1 trans-acting factors could also participate in the enhancing activity of this element. Taken together, the results demonstrate that two GC box-like sequences and another trans-acting factor-binding sequence located between -970 and -941 bp act coordinately and are essential for efficient transcription of this low expressed human ChAT gene.

Original languageEnglish
Pages (from-to)299-304
Number of pages6
JournalMolecular Brain Research
Volume20
Issue number4
DOIs
StatePublished - Dec 1993

Bibliographical note

Funding Information:
AJ~nowledgments We are gratetul to Dr R Sanders Wllhams for the genorous gift of the pGuP PA8 vector We also thank Dr Nono Kagawa lor helpful discussions during the progress of this work This work was supported in part by Grants AG05893 and AG08013 from the National Institutes of Health

Keywords

  • Choline acetyltransferase
  • Cis-acting factor
  • DNA-binding protein
  • Enhancer
  • GC box-like sequence
  • Sp1

ASJC Scopus subject areas

  • Molecular Biology
  • Cellular and Molecular Neuroscience

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