CHARACTERIZATION OF THE DNF15S2 LOCUS ON HUMAN-CHROMOSOME .3. IDENTIFICATION OF A GENE CODING FOR 4 KRINGLE DOMAINS WITH HOMOLOGY TO HEPATOCYTE GROWTH-FACTOR
CHARACTERIZATION OF THE DNF15S2 LOCUS ON HUMAN-CHROMOSOME .3. IDENTIFICATION OF A GENE CODING FOR 4 KRINGLE DOMAINS WITH HOMOLOGY TO HEPATOCYTE GROWTH-FACTOR
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DOI：
10.1021/bi00104a029
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发表时间：
1991-10-08
期刊：
影响因子：
2.9
通讯作者：
DEGEN, SJF
中科院分区：
文献类型：
--
作者：
HAN, S;STUART, LA;DEGEN, SJF
为了分离人凝血酶原基因，通过使用降低严格性的条件用编码凝血酶原中的kringle结构域的牛cDNA探针筛选人基因组DNA文库。 鉴定出十二个阳性结果，其中三个编码为凝血酶原（Degen & Davie，1987）。 由于噬菌体 L5 与 cDNA 探针的强杂交以及与编码人凝血酶原的基因相比其独特的限制性图谱，因此对噬菌体 L5 进行了更详细的表征。 L5 中的基因被测序并发现编码含有 kringle 的蛋白质。 使用L5基因的基因组探针筛选人肝脏cDNA文库。 分离出含有与L5基因中的区域相同的序列的cDNA。 cDNA与基因的比较表明，L5中的基因由18个外显子组成，中间有17个插入序列，长度为4690 bp。 外显子的长度范围为 36 至 242 bp，而插入序列的长度范围为 77 至 697 bp。 L5 中的基因编码的假定蛋白质包含四个 kringle 结构域，后面跟着一个丝氨酸蛋白酶样结构域。 该结构域结构与肝细胞生长因子 (HGF) 中的结构域相同，尽管这两种蛋白质只有约 50% 相同。 基于L5和HGF编码的蛋白质的相似性，我们建议在功能确定之前将推定的L5蛋白质暂时称为HGF样蛋白质。 将基因和cDNA的DNA序列及其翻译的氨基酸序列与GenBank和NBRF数据库进行比较。 在基因特征序列 3' 端的外显子 17 中鉴定出与 DNF15S1 和 DNF15S2、人 DNF15S2 肺 mRNA 和大鼠酰基肽水解酶同源的序列。 从我们的结果来看，很明显，编码人类 HGF 样蛋白的基因位于人类 3 号染色体 (3p21) 上的 DNF15S2 基因座。 酰基肽水解酶基因位于 HGF 样蛋白编码基因下游 444 bp，但位于互补链上。 DNF15S2 基因座已被提议编码一种或多种肿瘤抑制基因，因为该基因座在小细胞肺癌、其他肺癌、肾细胞癌和 von Hippel-Lindau 综合征的 DNA 中被删除。
A human genomic DNA library was screened by using conditions of reduced stringency with a bovine cDNA probe coding for the kringle domains in prothrombin in order to isolate the human prothrombin gene. Twelve positives were identified, three of which coded for prothrombin (Degen & Davie, 1987). Phage L5 was characterized in more detail because of its strong hybridization to the cDNA probe and its unique restriction map compared to the gene coding for human prothrombin. The gene in L5 was sequenced and found to code for a kringle-containing protein. A human liver cDNA library was screened by using a genomic probe from the gene in L5. cDNAs were isolated that contained sequence identical with regions in the gene in L5. Comparison of the cDNA with the gene indicated that the gene in L5 was composed of 18 exons separated by 17 intervening sequences and is 4690 bp in length. Exons ranged in size from 36 to 242 bp in length while intervening sequences ranged from 77 to 697 bp in length. The putative protein encoded by the gene in L5 contains four kringle domains followed by a serine protease-like domain. This domain structure is identical with that found in hepatocyte growth factor (HGF), although the two proteins are only about 50% identical. On the basis of the similarity of the protein encoded by L5 and HGF, we propose that the putative L5 protein be tentatively called HGF-like protein until a function is identified. The DNA sequence of the gene and cDNA and its translated amino acid sequence were compared against GenBank and NBRF databases. Sequences homologous to DNF15S1 and DNF15S2, human DNF15S2 lung mRNA, and rat acyl-peptide hydrolase were identified in exon 17 to the 3' end of the characterized sequence for the gene. From our results, it is apparent that the gene coding for human HGF-like protein is located at the DNF15S2 locus on human chromosome 3 (3p21). The gene for acyl-peptide hydrolase is 444 bp downstream of the gene coding for HGF-like protein, but on the complementary strand. The DNF15S2 locus has been proposed to code for one or more tumor suppressor genes since this locus is deleted in DNA from small cell lung carcinoma, other lung cancers, renal cell carcinoma, and von Hippel-Lindau syndrome.