BIOSYNTHESIS OF HUMAN FIBROBLAST GROWTH FACTOR-5
BIOSYNTHESIS OF HUMAN FIBROBLAST GROWTH FACTOR-5
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DOI：
10.1128/mcb.11.4.1840
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发表时间：
1991-04-01
影响因子：
5.3
通讯作者：
GOLDFARB, M
中科院分区：
文献类型：
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作者：
BATES, B;HARDIN, J;GOLDFARB, M
我们分析了人成纤维细胞生长因子-5（FGF-5）在翻译和翻译后水平的生物合成。 体外合成的FGF-5 RNA可以在兔网织红细胞裂解物中翻译，产生29，500-Da的蛋白质，这与从编码序列预测的分子量一致。 如果RNA中的上游开放阅读框（ORF-1）被缺失或如果ORF-1中的两个AUG密码子被点突变破坏，FGF-5翻译的效率显著增强，而通过ORF-1 AUG密码子的任一个的单独突变实现部分增强。 这些数据表明，FGF-5的合成需要扫描核糖体过去的两个ORF-1 AUG密码子。将这些ORF-1突变引入真核FGF-5表达载体后，其转化小鼠NIH 3 T3细胞的能力在转染后增加了50倍。 FGF-5从转染的3 T3细胞和人肿瘤细胞中分泌，作为含有不同量唾液酸的糖蛋白。 糖苷酶处理表明生长因子具有N-连接和O-连接的糖。
We have analyzed the biosynthesis of human fibroblast growth factor-5 (FGF-5) at the translational and posttranslational levels. FGF-5 RNA synthesized in vitro can be translated in rabbit reticulocyte lysates to yield a 29,500-Da protein, which is consistent with the molecular weight predicted from the coding sequence. The efficiency of FGF-5 translation is dramatically enhanced if an upstream open reading frame (ORF-1) in the RNA is deleted or if both AUG codons in ORF-1 are destroyed by point mutations, while partial enhancement is achieved by individual mutation of either ORF-1 AUG codon. These data suggest that FGF-5 synthesis requires the scanning of ribosomes past the two ORF-1 AUG codons. The introduction of these ORF-1 mutations into a eukaryotic FGF-5 expression vector increases its capacity to transform mouse NIH 3T3 cells up to 50-fold upon transfection. FGF-5 is secreted from transfected 3T3 cells and from human tumor cells as glycoproteins containing heterogeneous amounts of sialic acid. Glycosidase treatments suggest that the growth factor bears both N-linked and O-linked sugars.