Regulation of vimentin gene transcription in human breast cancer cell lines.
We have investigated the control of vimentin expression in human breast cancer cell lines because of its transcriptional activation during malignant progression in breast cancer. Comparison of vimentin-positive (V+) and vimentin-negative (V-) breast cancer cell lines revealed several potential areas of vimentin gene regulation. Analysis of the chromatin structure of the vimentin gene in V+ and V- breast cancer cells showed DNase I hypersensitive sites in the 5' promoter region in V+ cell lines and 3' to the start of transcription in V- cell lines. Promoter deletion and reporter gene analysis revealed the importance of two adjacent AP-1 sites separated by seven GC-rich nucleotides for vimentin expression in V+ breast cancer cells. Mutational analysis of these sequences showed that although both AP-1 sites could bind nuclear proteins from V+ cells in vitro, one AP-1 site was sufficient to drive transcription in CAT reporter gene assays. The GC-rich spacer region had a modulating function on the activity of the AP-1 sites. In addition, levels of c-jun mRNA were elevated in V+ versus V- cells. In summary, distinct sites within the vimentin gene appear to be important for the control of vimentin expression in V+ and V- breast cancer cells with multiple elements acting coordinately to regulate vimentin expression.
Sommers
C L
CL
Department of Medicine, Georgetown University School of Medicine, Washington, DC 20007.
Skerker
J M
JM
Chrysogelos
S A
SA
Bosseler
M
M
Gelmann
E P
EP
D001483:Base Sequence; D001943:Breast Neoplasms / Q000235:genetics* / Q000473:pathology; D002467:Cell Nucleus; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics*; D003850:Deoxyribonuclease I; D017930:Genes, Reporter; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D018808:Transcription Factor AP-1 / Q000235:genetics; D014158:Transcription, Genetic / Q000502:physiology*; D014407:Tumor Cells, Cultured; D014746:Vimentin / Q000235:genetics*
D018808:Transcription Factor AP-1; D014746:Vimentin; D015500:Chloramphenicol O-Acetyltransferase; D003850:Deoxyribonuclease I