Regulation of the calcitonin/calcitonin gene-related peptide gene by cell-specific synergy between helix-loop-helix and octamer-binding transcription factors

Lois A Tverberg; Andrew F Russo

doi:10.1016/S0021-9258(18)82346-8

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Regulation of the calcitonin/calcitonin gene-related peptide gene by cell-specific synergy between helix-loop-helix and octamer-binding transcription factors

Journal article

Open access

Peer reviewed

Regulation of the calcitonin/calcitonin gene-related peptide gene by cell-specific synergy between helix-loop-helix and octamer-binding transcription factors

Lois A Tverberg and Andrew F Russo

The Journal of biological chemistry, Vol.268(21), pp.15965-15973

07/25/1993

DOI: 10.1016/S0021-9258(18)82346-8

PMID: 8340417

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Abstract

The calcitonin/calcitonin gene-related peptide (CGRP) gene is transcribed in thyroid C-cells and a subset of neurons. We have localized sequences required for cell-specific enhancement of calcitonin/CGRP transcription in rat thyroid C-cell lines. An 18-base pair element approximately 1 kilobase pair upstream of the transcriptional start site stimulated expression of a luciferase reporter gene 50-fold in 44-2C C-cells. There was less than 2-fold stimulation in HeLa and Rat-1 cells, which do not express the endogenous calcitonin/CGRP gene. The enhancer contains potential binding sites for helix-loop-helix (HLH) and octamer transcription factors based on sequence homologies. The functional significance of these sites was shown by point mutations in the HLH and octamer motifs and by separation of the two motifs, all of which decreased enhancer activity greater than 10-fold. The involvement of HLH proteins was further shown by co-expression of the mammalian achaete-scute homologue-1 HLH protein, which activated the enhancer severalfold in HeLa cells. Electrophoretic mobility shift analyses revealed several DNA-protein complexes containing HLH and octamer-binding proteins. One octamer-binding complex (OB1) most likely contains the ubiquitous Oct-1 protein, whereas a second complex (OB2) was cell-specific. In contrast to OB1, OB2 had lower affinity for a consensus octamer motif, and its DNA binding was not affected by addition of antiserum that recognizes Oct-1 and Oct-2 proteins. In addition, we observed a large complex that appears to contain both an HLH protein and OB2. These results demonstrate that calcitonin/CGRP enhancer activity is controlled by a cell-specific synergistic activation involving HLH and octamer-binding factors.

Binding, Competitive

Cell Line

Octamer Transcription Factor-2

DNA-Binding Proteins - immunology

Octamer Transcription Factor-1

Immune Sera

Humans

Gene Expression Regulation

Molecular Sequence Data

Rats

Calcitonin Gene-Related Peptide - genetics

DNA - metabolism

DNA-Binding Proteins - metabolism

Transcription Factors - metabolism

Animals

Enhancer Elements, Genetic

Host Cell Factor C1

Base Sequence

HeLa Cells

Calcitonin Gene-Related Peptide - metabolism

Details

Title: Subtitle: Regulation of the calcitonin/calcitonin gene-related peptide gene by cell-specific synergy between helix-loop-helix and octamer-binding transcription factors
Creators: Lois A Tverberg - Department of Physiology and Biophysics, University of Iowa, Iowa City 52242
Andrew F Russo
Resource Type: Journal article
Publication Details: The Journal of biological chemistry, Vol.268(21), pp.15965-15973
DOI: 10.1016/S0021-9258(18)82346-8
PMID: 8340417
NLM abbreviation: J Biol Chem
ISSN: 0021-9258
eISSN: 1083-351X
Publisher: United States
Grant note: HD25969 / NICHD NIH HHS DK25295 / NIDDK NIH HHS
Language: English
Date published: 07/25/1993
Academic Unit: Neurology; Molecular Physiology and Biophysics; Iowa Neuroscience Institute; Craniofacial Anomalies Research Center
Record Identifier: 9984020869202771

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