Cerretti D P, Wignall J, Anderson D, Tushinski R J, Gallis B M, Stya M, Gillis S, Urdal D L, Cosman D
Department of Molecular Biology, Immunex Corporation, Seattle, Washington 98101.
Mol Immunol. 1988 Aug;25(8):761-70. doi: 10.1016/0161-5890(88)90112-5.
Macrophage-colony stimulating factor (M-CSF, CSF-1) has been reported to be required for the proliferation and differentiation of macrophages from hematopoietic progenitor cells. Recently, two human M-CSF cDNA clones were isolated encoding proteins of 256 and 554 amino acids. We report here the isolation of a third M-CSF cDNA that encodes a protein of 438 amino acids. The coding regions for the three cDNA clones share a common amino-terminus of 149 amino acids and a common carboxyl-terminus of 75 amino acids including a membrane spanning region. In addition, we isolated a genomic clone of human M-CSF. When each of the cDNA clones or the genomic clone were transfected into COS-7 monkey kidney cells, biologically active M-CSF was expressed as judged by the ability of transfected cell supernatants to stimulate proliferation and colony formation of murine bone marrow cells, as well as formation of monocytic colonies from human bone marrow cells. Surprisingly, proliferation of human bone marrow cells was not induced by recombinant human M-CSF. Analysis of the M-CSF proteins released by COS-7 cells revealed that monomer subunit proteins of 44 or 28 kDa were produced. In addition, we found that the membrane spanning region, present in all three forms of M-CSF cDNA, was not required for the synthesis of a biologically active protein. However, when the membrane spanning region was present in the three M-CSF cDNAs, cell surface associated forms of M-CSF could be readily detected.
据报道，巨噬细胞集落刺激因子（M-CSF，CSF-1）是造血祖细胞分化为巨噬细胞所必需的。最近，分离出了两个编码256和554个氨基酸的人M-CSF cDNA克隆。我们在此报告分离出了第三个M-CSF cDNA，其编码一种438个氨基酸的蛋白质。这三个cDNA克隆的编码区共有一个149个氨基酸的共同氨基末端和一个75个氨基酸的共同羧基末端，包括一个跨膜区。此外，我们还分离出了人M-CSF的基因组克隆。当将每个cDNA克隆或基因组克隆转染到COS-7猴肾细胞中时，通过转染细胞上清液刺激小鼠骨髓细胞增殖和集落形成以及人骨髓细胞形成单核细胞集落的能力判断，可表达出具有生物活性的M-CSF。令人惊讶的是，重组人M-CSF并未诱导人骨髓细胞的增殖。对COS-7细胞释放的M-CSF蛋白的分析表明，产生了44或28 kDa的单体亚基蛋白。此外，我们发现，在所有三种形式的M-CSF cDNA中都存在的跨膜区，对于生物活性蛋白的合成并非必需。然而，当跨膜区存在于三种M-CSF cDNA中时，很容易检测到细胞表面相关形式的M-CSF。
