Tannenbaum C S, Major J, Ohmori Y, Hamilton T A
Research Institute, Cleveland Clinic Foundation, Ohio 44195.
J Leukoc Biol. 1993 May;53(5):563-8. doi: 10.1002/jlb.53.5.563.
We previously reported the isolation and characterization of cDNA clones encoding novel lipopolysaccharide (LPS)-inducible mRNAs from murine peritoneal macrophages. We now present the complete coding sequence of a cDNA previously termed D3. Analysis of multiple clones from a murine macrophage cDNA library provided a complete cDNA sequence of approximately 1.6 kb. The corresponding RNA contains a single open reading frame encoding a hydrophilic protein composed of 425 amino acids and is characterized by a region including three perfect and two imperfect repeats of a seven-amino-acid sequence. Based on nucleotide and deduced amino acid sequence, this mRNA is a new member of a previously described multigene cluster of interferon-inducible genes termed the Mouse 200 series genes. This new sequence most closely resembles gene 204 because both D3 and 204 genes have segments containing the seven-amino-acid repeat sequence. The Mouse 202 and 204 genes, however, have an approximately 200-amino-acid carboxyl-terminal domain that is absent in the LPS-inducible macrophage-derived cDNA. In addition, D3, 202, and 204 can all be distinguished from one another by virtue of unique 3' noncoding regions 200-300 base pairs in length. The D3 unique sequence is largely restricted to the smallest of the three size classes of this gene family expressed in macrophages and is not detected in interferon- or platelet-derived growth factor-stimulated fibroblasts. Overall, three separate mRNAs have now been described, each of which has three or more of a possible seven nucleotide sequence domains. Although the function(s) of the members of this gene family remains unknown, the multiple forms inducible by diverse stimuli and their restricted cell type expression suggest diverse and important physiologic roles for their products in inflammation.
我们之前报道了从鼠腹膜巨噬细胞中分离和鉴定编码新型脂多糖(LPS)诱导型mRNA的cDNA克隆。现在我们展示了一个之前称为D3的cDNA的完整编码序列。对鼠巨噬细胞cDNA文库中多个克隆的分析提供了一个约1.6 kb的完整cDNA序列。相应的RNA包含一个单一的开放阅读框,编码一个由425个氨基酸组成的亲水蛋白,其特征是一个区域包含一个七氨基酸序列的三个完美重复和两个不完美重复。基于核苷酸和推导的氨基酸序列,该mRNA是先前描述的干扰素诱导基因多基因簇(称为小鼠200系列基因)的一个新成员。这个新序列与基因204最相似,因为D3和204基因都有包含七氨基酸重复序列的片段。然而,小鼠202和204基因有一个约200个氨基酸的羧基末端结构域,而在LPS诱导的巨噬细胞来源的cDNA中不存在。此外,D3、202和204都可以通过长度为200 - 300个碱基对的独特3'非编码区相互区分。D3独特序列主要局限于巨噬细胞中表达的该基因家族三个大小类别中最小的那个类别,在干扰素或血小板衍生生长因子刺激的成纤维细胞中未检测到。总体而言,现在已经描述了三种不同的mRNA,每种都有三个或更多可能的七核苷酸序列结构域。虽然这个基因家族成员的功能仍然未知,但多种形式可被不同刺激诱导以及它们受限的细胞类型表达表明其产物在炎症中具有多样且重要的生理作用。