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Heregulin and forskolin‐induced cyclin D3 expression in Schwann cells: Role of a CCAAT promoter element and CCAAT enhancer binding protein
Author(s) -
Fuentealba Luis,
Schworer Charles,
Schroering Allen,
Rahmatullah Mohammed,
Carey David J.
Publication year - 2004
Publication title -
glia
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.954
H-Index - 164
eISSN - 1098-1136
pISSN - 0894-1491
DOI - 10.1002/glia.10325
Subject(s) - forskolin , biology , microbiology and biotechnology , caat box , neuregulin , ccaat enhancer binding proteins , reporter gene , transcription factor , promoter , gene expression , signal transduction , cell culture , nuclear protein , gene , genetics
Heregulin, a polypeptide growth factor, and forskolin, an adenylyl cyclase activator, synergistically stimulate expression of cyclin D3 and cell division in Schwann cells. Heregulin induces expression in Schwann cells of a luciferase reporter gene linked to the cyclin D3 promoter. Forskolin markedly augments reporter expression in the presence of heregulin. Deletion analysis identified several promoter sites that contribute to high‐level reporter expression in heregulin‐ and forskolin‐treated Schwann cells. A promoter fragment that contains 103 bp of 5′‐flanking sequence produced significant reporter expression in heregulin‐ and forskolin‐stimulated cells. Deletion of a consensus CCAAT site within this promoter fragment caused a nearly complete loss of reporter expression. Similar results were obtained when CCAAT site mutations were introduced into the promoter. Heregulin and forskolin increased steady‐state levels of CCAAT/enhancer binding protein‐β (C/EBPβ) in Schwann cells. Mobility shift assays identified proteins in Schwann cell nuclear extracts that formed stable complexes with the cyclin D3 CCAAT promoter element and were disrupted by anti‐C/EBPβ antibody. Transfection of Schwann cells with C/EBPβ cDNA increased cyclin D3 reporter expression. In contrast to these results, mutation of a cAMP response element in the cyclin D3 promoter had only a modest effect on heregulin‐ and forskolin‐stimulated reporter expression. These findings demonstrate that C/EBPβ plays a key role in the heregulin and cAMP‐dependent regulation of cyclin D3 expression in Schwann cells. © 2003 Wiley‐Liss, Inc.