Farber J M
Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205.
Mol Cell Biol. 1992 Apr;12(4):1535-45. doi: 10.1128/mcb.12.4.1535-1545.1992.
To identify genes induced during macrophage activation, a cDNA library was prepared from cultures of the RAW 264.7 mouse macrophage cell line that had been treated with conditioned medium from mitogen-stimulated spleen cells, and the cDNA library was screened by differential plaque hybridization. Eleven cDNA clones, designated CRG-1 through CRG-11, corresponding to mRNA species inducible in RAW 264.7 cells by the spleen cell conditioned medium, were isolated. Inductions were not blocked by cycloheximide. All of the mRNAs were inducible by gamma interferon, and some were also inducible by alpha and beta interferons, by lipopolysaccharide, by phorbol 12-myristate 13-acetate, and by the calcium ionophore A23187. Sequencing of the cDNAs revealed that CRG-1, CRG-3, and CRG-5 are cDNAs of recently identified transcription factors IRF-1, zif/268, and LRF-1 respectively. As previously reported, CRG-2 and CRG-10 (MIG) encode new members of the platelet factor 4 family of cytokines. CRG-6 corresponds to a new member of a family of interferon-inducible genes clustered on mouse chromosome 1, CRG-9 corresponds to a prostaglandin synthase homolog, CRG-8 corresponds to beta 2-microglobulin, and CRG-4 corresponds to metallothionein II. CRG-11 contains sequences of a truncated L1Md repetitive element as well as nonrepetitive sequences. The nonrepetitive sequence of CRG-11 as well as the sequences of CRG-7 are not closely related to published sequences. The CRG genes and proteins are of interest because of their involvement in macrophage activation, because of their roles as mediators of the effects of gamma interferon and other pleiotropic agents, and because of their usefulness as tools for studying the signal pathways through which gamma interferon and other inducers exert their effects on gene and protein expression.
为了鉴定巨噬细胞激活过程中诱导表达的基因,从经丝裂原刺激的脾细胞条件培养基处理过的RAW 264.7小鼠巨噬细胞系培养物中制备了一个cDNA文库,并通过差异噬菌斑杂交对该cDNA文库进行筛选。分离出了11个cDNA克隆,命名为CRG-1至CRG-11,它们对应于RAW 264.7细胞中可被脾细胞条件培养基诱导的mRNA种类。诱导作用不受放线菌酮的阻断。所有这些mRNA均可被γ干扰素诱导,有些还可被α和β干扰素、脂多糖、佛波酯12-肉豆蔻酸酯13-乙酸酯以及钙离子载体A23187诱导。对这些cDNA进行测序发现,CRG-1、CRG-3和CRG-5分别是最近鉴定出的转录因子IRF-1、zif/268和LRF-1的cDNA。如先前报道的那样,CRG-2和CRG-10(MIG)编码血小板因子4细胞因子家族的新成员。CRG-6对应于小鼠1号染色体上成簇的干扰素诱导基因家族的一个新成员,CRG-9对应于一种前列腺素合酶同源物,CRG-8对应于β2-微球蛋白,CRG-4对应于金属硫蛋白II。CRG-11包含一个截短的L1Md重复元件序列以及非重复序列。CRG-11的非重复序列以及CRG-7的序列与已发表的序列没有密切关系。CRG基因和蛋白质之所以受到关注,是因为它们参与巨噬细胞激活,作为γ干扰素和其他多效性因子作用的介质,以及作为研究γ干扰素和其他诱导剂对基因和蛋白质表达发挥作用的信号通路的工具。