Abstract
The addition of polypeptide mitogens to quiescent cell lines induces the expression of various gene products, some of which are likely to perform functions critical for cell cycle progression, DNA synthesis, and mitosis. We have used a differential display approach to identify fibroblast growth factor (FGF)-1-inducible genes in NIH-3T3 cells. One of these genes, termed FGF-regulated (FR)-1, encodes a 316-amino acid protein with approximately 82% amino acid sequence identity to an abundant protein expressed in mouse vas deferens and approximately 70% identity to human aldose reductase. The function of the vas deferens protein is unknown; however, aldose reductase is an NADPH-dependent monomeric oxidoreductase implicated in the pathogenesis of diabetic complications. FGF-1 induction of FR-1 mRNA expression is first detectable at 4 h after mitogen addition and is dependent on de novo RNA and protein synthesis. FGF-2 or phorbol ester treatment can also increase FR-1 mRNA levels; in contrast, whole blood serum or individual growth factors present in serum have only minimal effects on FR-1 mRNA expression. FR-1 mRNA is detectable in a number of mouse tissues but is most abundant in newborn liver and in adult intestine, ovary, and testis. These results raise the possibility that aldose reductase-related proteins may play a role in FGF-1- and FGF-2-stimulated mitogenesis.
Highlights
The addition of polypeptide mitogensto quiescent cell thoughitisknown that fibroblast growth factor (FGF)-1bindinginitiatesnumerous lines induces the expression of various gene products, biochemical changes, including the activation of specific genes some of which arelikely to perform functionscriticalfor [1]
We present data inof mouse tissues but is most abundant in newbornliver dicating that FGF-1 can inducethe expression of a transcript and in adult intestine, ovary, and testis
The gels were stained with ethidium bromide to could regulate FR-1 mRNA levels in serum-starved NIH 3T3 verify that each lane contained similar amountsof undegraded rRNA. cells
Summary
Dept. of Molecular Biology, Holland Laboratory, American Red Cross, 15601 Crabbs Branch Way, Rockville, MD 20855.Tel.:301-738-0655. InitiaRl NA gel blot hybridization experiments indicated that thisDNA fragment hybridized to a transcript of -1.5 kilobases in size This fragment,termed FR-1, was radiolabeled and used toscreen a DNA polymerase (Boehringer Mannheim), 0.25 pgof a zinc finger do- Balb/c 3T3 cell hgtl0 cDNA library in order to isolate larger main sense primer, and0.25 pg of a tyrosine kinase domain antisense primer. The gels were stained with ethidium bromide to could regulate FR-1 mRNA levels in serum-starved NIH 3T3 verify that each lane contained similar amountsof undegraded rRNA. The FR-1mRNA signal was normalized gene expression In this particular experiment, FR-1 mRNA levels were increased -11.5-fold after 8 h of FGF-1 treatment (Fig. 2 B ). Serum itself decreased FR-1 mRNA levels, some of the major growth
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