Miyazaki Y, Setoguchi M, Yoshida S, Higuchi Y, Akizuki S, Yamamoto S
Department of Pathology, Medical College of Oita, Japan.
J Biol Chem. 1990 Aug 25;265(24):14432-8.
Murine macrophage cell lines and resident macrophages showed various levels of expression of the murine osteopontin (OP) gene, and macrophage stimulating agents were found to enhance transcription of the gene with kinetics which are unique for each stimulator. The organization of the murine OP gene was determined. The gene comprises six exons and five introns and spans approximately 4.8 kilobases. Exon 1 contains the 16 amino acids of the leader sequence. Exons 2, 3, 4, 5, and 6 encode 12, 27, 14, 94, and 129 amino acid residues, respectively. Exon 5 encodes regions containing 10 consecutive Asp amino acid residues and a Gly-Arg-Gly-Asp-Ser peptide. Exon 6 encodes the C-terminal half of OP and contains no 15- and 54-base pair nucleotide sequences which are deleted in murine OP cDNA compared to that of rat OP cDNA. Since Southern blot analysis indicated that the OP gene is a single copy, it is obvious that the murine OP cDNA has the sequence previously determined (Miyazaki, Y., Setoguchi, M., Yoshida, S., Higuchi, Y., Akizuki, S., and Yamamoto, S. (1989) Nucleic Acids Res. 17, 3298). A comparison with the cDNA sequences reported previously suggested the presence of nucleotide sequence polymorphisms. The 5' end of the murine OP gene was defined by primer extension and S1 nuclease mapping. Sequence analysis of the 5'-flanking DNA revealed the presence of many potential regulatory motifs.
小鼠巨噬细胞系和驻留巨噬细胞显示出不同水平的小鼠骨桥蛋白(OP)基因表达,并且发现巨噬细胞刺激剂能够增强该基因的转录,其动力学特征因每种刺激剂而异。确定了小鼠OP基因的结构。该基因由六个外显子和五个内含子组成,跨度约为4.8千碱基。外显子1包含前导序列的16个氨基酸。外显子2、3、4、5和6分别编码12、27、14、94和129个氨基酸残基。外显子5编码包含10个连续天冬氨酸氨基酸残基和一个甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸肽的区域。外显子6编码OP的C端一半,并且与大鼠OP cDNA相比,在小鼠OP cDNA中不存在缺失的15和54碱基对核苷酸序列。由于Southern印迹分析表明OP基因是单拷贝的,显然小鼠OP cDNA具有先前确定的序列(宫崎洋、濑户口正、吉田秀、樋口洋、秋津树和山本幸男(1989年)《核酸研究》17,3298)。与先前报道的cDNA序列比较表明存在核苷酸序列多态性。通过引物延伸和S1核酸酶图谱确定了小鼠OP基因的5'端。对5'侧翼DNA的序列分析揭示了许多潜在的调控基序的存在。