Heard J M, Roussel M F, Rettenmier C W, Sherr C J
Department of Tumor Cell Biology, St. Jude Children's Research Hospital, Memphis, TN 38105.
Oncogene Res. 1987 Sep-Oct;1(4):423-40.
The mononuclear phagocyte colony stimulating factor encoded by a 1.6 kilobase pair human cDNA is synthesized as a homodimeric transmembrane glycoprotein that is released from the plasma membrane by proteolysis. Premature termination of the CSF-1 coding sequence upstream of its carboxylterminal transmembrane-spanning segment and expression of the truncated CSF-1 cDNA in either bovine papilloma virus or retrovirus vectors led to the synthesis of a soluble, biologically active growth factor that was rapidly secreted from cells. Like the full-length CSF-1 precursor, the truncated polypeptide was rapidly assembled through disulfide bonds immediately after synthesis and acquired asparagine-linked oligosaccharide chains that underwent progressive post-translational modifications during intracellular transport. Soluble CSF-1 encoded by the truncated cDNA stimulated the formation of bone marrow-derived mouse macrophage colonies in semisolid medium and induced transformation of mouse NIH-3T3 cells when coexpressed with the human c-fms proto-oncogene product (CSF-1 receptor). Compared to results obtained with the full-length CSF-1 cDNA, the efficiency of transformation obtained with the truncated CSF-1 gene was reduced, in spite of the fact that transfected cultures produced similar levels of the extracellular growth factor. The results indicate that CSF-1 amino acid residues 1-158 (together with the aminoterminal signal peptide at residues -32 to -1) are sufficient for biological activity and that CSF-1 cDNAs encoding either membrane-bound or soluble precursors are active in autocrine transformation.
由一个1.6千碱基对的人cDNA编码的单核吞噬细胞集落刺激因子被合成为一种同二聚体跨膜糖蛋白,该蛋白通过蛋白水解作用从质膜释放。在其羧基末端跨膜片段上游的CSF-1编码序列提前终止,并在牛乳头瘤病毒或逆转录病毒载体中表达截短的CSF-1 cDNA,导致合成一种可溶性的、具有生物活性的生长因子,该因子能迅速从细胞中分泌出来。与全长CSF-1前体一样,截短的多肽在合成后立即通过二硫键迅速组装,并获得天冬酰胺连接的寡糖链,这些寡糖链在细胞内运输过程中经历了渐进的翻译后修饰。截短的cDNA编码的可溶性CSF-1在半固体培养基中刺激骨髓来源的小鼠巨噬细胞集落的形成,当与人c-fms原癌基因产物(CSF-1受体)共表达时,可诱导小鼠NIH-3T3细胞的转化。与全长CSF-1 cDNA的结果相比,尽管转染培养物产生的细胞外生长因子水平相似,但截短的CSF-1基因的转化效率降低。结果表明,CSF-1氨基酸残基1-158(连同残基-32至-1处的氨基末端信号肽)足以产生生物活性,并且编码膜结合或可溶性前体的CSF-1 cDNA在自分泌转化中具有活性。