TY - JOUR
T1 - Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA
AU - Payson, Robert A.
AU - Canatan, Halit
AU - Chotani, Maqsood A.
AU - Wang, Wen pin
AU - Harris, Stephen E.
AU - Myers, René L.
AU - Chiu, Ing ming
N1 - Funding Information:
We thank Julia Wilson for her assistance in preparing the manuscript and Runny Bergman for her excellent technical assistance. This work was supported in part by grants from the National Cancer Institute (R01 CA45611) and the Ohio Cancer Research Associates, Inc. I.-M.C. is a recipient of the Research Career Development Award (K04 CA01369) from the National Institutes of Health. R.A.P. and R.L.M. are supported by NIH NRSA T32 CA09498 and T32 CAO9338, respectively. H.C. is a recipient of a NATO Science fellowship via the Scientific and Technical Research Council of Turkey.
PY - 1993/2/11
Y1 - 1993/2/11
N2 - We have previously isolated two different aFGF cDNA clones from kidney and brain. The two corresponding mRNA, designated aFGF 1.A and 1.B, are the predominant species in kidney and brain, respectively. During the characterization of aFGF mRNA in glioblastoma ceils, we demonstrated that aFGF mRNA in U1242MG and D65MG gliobiastoma cells contain 5′-untransiated sequences different from those of 1 .A and 1.B. Through a strategy combining chromosome wafldng, Identification and sequencing of evolutlonarily conserved DNA regions, and a reverse transcription and polymerase chain reaction (RT-PCR)-based assay for RNA expression, we have isoiated two novel aFGF cDNA clones. The cDNA clone representing aFGF mRNA 1.C was isolated from U1242MG cells; another aFGF cDNA, designated 1.D, was isolated from D65MG cells. Promoter 1C has extensive sequence homology to the hamster aFGF gene promoter that was shown to respond to testosterone stimulation by chioramphenicol acetyltransferase reporter gene assays. Using RT-PCR, we showed that normal, benign and cancerous prostate tissues do not express aFGF 1.C mRNA. in contrast, a prostate carcinoma cell line (PC-3) expresses 1.C mRNA. RT-PCR using 1.D-specific primers showed that kidney, brain and prostate do not express 1.D mRNA even though kidney and brain are the most abundant source for aFGF protein. RNase protection analysis further showed that 1.D mRNA is the predominant aFGF transcript In D65MG giiobiastoma cells and in NFF-6 neonatal foreskin fibrobiast cells. The genomic DNA corresponding to these two cDNA clones and the 5′-fianklng regions were also isolated and their sequences determined. These DNA clones will provide important reagents for studying the regulatory elements of aFGF gene expression.
AB - We have previously isolated two different aFGF cDNA clones from kidney and brain. The two corresponding mRNA, designated aFGF 1.A and 1.B, are the predominant species in kidney and brain, respectively. During the characterization of aFGF mRNA in glioblastoma ceils, we demonstrated that aFGF mRNA in U1242MG and D65MG gliobiastoma cells contain 5′-untransiated sequences different from those of 1 .A and 1.B. Through a strategy combining chromosome wafldng, Identification and sequencing of evolutlonarily conserved DNA regions, and a reverse transcription and polymerase chain reaction (RT-PCR)-based assay for RNA expression, we have isoiated two novel aFGF cDNA clones. The cDNA clone representing aFGF mRNA 1.C was isolated from U1242MG cells; another aFGF cDNA, designated 1.D, was isolated from D65MG cells. Promoter 1C has extensive sequence homology to the hamster aFGF gene promoter that was shown to respond to testosterone stimulation by chioramphenicol acetyltransferase reporter gene assays. Using RT-PCR, we showed that normal, benign and cancerous prostate tissues do not express aFGF 1.C mRNA. in contrast, a prostate carcinoma cell line (PC-3) expresses 1.C mRNA. RT-PCR using 1.D-specific primers showed that kidney, brain and prostate do not express 1.D mRNA even though kidney and brain are the most abundant source for aFGF protein. RNase protection analysis further showed that 1.D mRNA is the predominant aFGF transcript In D65MG giiobiastoma cells and in NFF-6 neonatal foreskin fibrobiast cells. The genomic DNA corresponding to these two cDNA clones and the 5′-fianklng regions were also isolated and their sequences determined. These DNA clones will provide important reagents for studying the regulatory elements of aFGF gene expression.
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U2 - 10.1093/nar/21.3.489
DO - 10.1093/nar/21.3.489
M3 - Article
C2 - 7680120
AN - SCOPUS:0027193268
SN - 0305-1048
VL - 21
SP - 489
EP - 495
JO - Nucleic acids research
JF - Nucleic acids research
IS - 3
ER -