Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA

Robert A. Payson, Halit Canatan, Maqsood A. Chotani, Wen Pin Wang, Stephen E Harris, René L. Myers, Ing Ming Chiu

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

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 cells, we demonstrated that aFGF mRNA in U1242MG and D65MG glioblastoma cells contain 5′-untranslated sequences different from those of 1 .A and 1.B. Through a strategy combining chromosome walking, identification and sequencing of evolutionarily conserved DNA regions, and a reverse transcription and polymerase chain reaction (RT-PCR)-based assay for RNA expression, we have isolated 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 chloramphenicol 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 glioblastoma cells and in NFF-6 neonatal foreskin fibroblast cells. The genomic DNA corresponding to these two cDNA clones and the 5′-flanking regions were also isolated and their sequences determined. These DNA clones will provide important reagents for studying the regulatory elements of aFGF gene expression.

Original languageEnglish (US)
Pages (from-to)489-495
Number of pages7
JournalNucleic Acids Research
Volume21
Issue number3
StatePublished - 1993

Fingerprint

Fibroblast Growth Factor 1
Fibroblasts
Cloning
Growth Factors
Messenger RNA
Organism Cloning
CDNA
Clone
Kidney
Cell
Polymerase Chain Reaction
Complementary DNA
Polymerase chain reaction
Brain
Clone Cells
Transcription
Glioblastoma
Reverse
DNA
Express

ASJC Scopus subject areas

  • Genetics
  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)

Cite this

Payson, R. A., Canatan, H., Chotani, M. A., Wang, W. P., Harris, S. E., Myers, R. L., & Chiu, I. M. (1993). Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA. Nucleic Acids Research, 21(3), 489-495.

Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA. / Payson, Robert A.; Canatan, Halit; Chotani, Maqsood A.; Wang, Wen Pin; Harris, Stephen E; Myers, René L.; Chiu, Ing Ming.

In: Nucleic Acids Research, Vol. 21, No. 3, 1993, p. 489-495.

Research output: Contribution to journalArticle

Payson, RA, Canatan, H, Chotani, MA, Wang, WP, Harris, SE, Myers, RL & Chiu, IM 1993, 'Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA', Nucleic Acids Research, vol. 21, no. 3, pp. 489-495.
Payson RA, Canatan H, Chotani MA, Wang WP, Harris SE, Myers RL et al. Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA. Nucleic Acids Research. 1993;21(3):489-495.
Payson, Robert A. ; Canatan, Halit ; Chotani, Maqsood A. ; Wang, Wen Pin ; Harris, Stephen E ; Myers, René L. ; Chiu, Ing Ming. / Cloning of two novel forms of human acidic fibroblast growth factor (aFGF) mRNA. In: Nucleic Acids Research. 1993 ; Vol. 21, No. 3. pp. 489-495.
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