Biochemical and Biophysical Research Communications
Genomic structure and promoter activity of the E1AF gene, a member of the ETS oncogene family
Section snippets
Materials and methods
Cells. Human prostatic adenocarcinoma PC3, human neuroblastoma GOTO, human oral squamous-cell carcinoma SAS, human embryo lung fibroblast MRC-5, and mouse mammary carcinoma FM3A (Health Science Research Resources Bank) were cultured at 37 °C with 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) with 5% fetal bovine serum and penicillin/streptomycin.
Isolation of genomic clones. A human genomic library of human peripheral blood cells (from Japanese Cancer Research Resources Bank, Tokyo, Japan)
Genome organization of the human E1AF gene
To obtain a genomic clone of E1AF, we screened a genomic library of human peripheral blood cells and a P1 phage system using primers corresponding to the 5′-end of E1AF cDNA [2] (see Materials and methods). The E1AF gene spanned approximately 30 kb and contained 14 exons as shown previously [17]. To clarify the start site, we carried out 5′-RACE (rapid amplification of the cDNA end) analysis using several cell lines such as PC3, SAS, MRC-5, and GOTO. Sequence analysis by comparing a nucleotide
Acknowledgments
We thank Dr. T. Matsuda for the expression constructs of estrogen receptor and Dr. Adachi for the genomic library. This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.
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2012, Biochimica et Biophysica Acta - Reviews on CancerCitation Excerpt :Together with the observed upregulation of PEA3 factors in breast cancer, these data implicate that PEA3 factor overexpression contributes to the causation of mammary tumors. Molecular studies suggest a vicious cycle: HER2/Neu activates the transcriptional activity of PEA3 factors [51,52], which in turn bind to the ETV4 and HER2/Neu gene promoters and stimulate their transcription [16,40,53]. This is controversial, since one report posits that ETV4 binds to and thereby represses the HER2/Neu gene promoter [54].
Adenovirus E1A negatively regulates E1AF, an ets family of the protein
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These two authors contributed equally to this work.
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Present address: Division of Protein Information, Institute for Genome Research, The University of Tokushima, 3-18-15, Kuramoto-cho 770-8503, Tokushima-city, Japan.