青石斑鱼抗细胞凋亡因子DAD1的克隆及表达特征分析

王利; 吴信忠

青石斑鱼抗细胞凋亡因子DAD1的克隆及表达特征分析

王利,
吴信忠

浙江大学, 动物科学学院, 浙江, 杭州, 310029

基金项目: 国家自然科学基金资助项目(30371107);浙江省重点科技计划项目(2004C23041)

计量
- 文章访问数: 634
- HTML全文浏览量: 9
- PDF下载量: 898
- 被引次数: 0
出版历程
- 收稿日期: 2006-07-17
- 修回日期: 2007-01-12

Molecular cloning and characterization of a homologue of defender against cell death 1 in Epinephelus awoara

College of Animal Sciences, Zhejiang University, Hangzhou 310029, China

摘要

摘要: 抗细胞凋亡因子(Defender against cell death 1,DAD1)是一种重要的内源性细胞凋亡抑制因子.采用RACE-PCR方法,结合SMART cDNA合成方法、抑制性差减杂交技术,从青石斑鱼脾脏中克隆DAD1并研究其在正常的青石斑鱼和注射了脂多糖的青石斑鱼各组织中的表达模式.结果表明:DAD1基因cDNA全长721 bp,包含一个342 bp的完整阅读框,编码一个由113个氨基酸组成的蛋白.RT-PCR半定量分析显示DAD1在正常的青石斑鱼和注射了脂多糖的青石斑鱼的脾、心、头肾、肾、肝中都有转录,在脾、肝中表达水平较高,而在肌肉中未见明显表达.DAD1在注射了脂多糖的青石斑鱼各组织中的表达量均高于正常的青石斑鱼.
- 青石斑鱼 /
- 抗细胞凋亡因子 /
- 克隆 /
- 表达
Abstract: Defender against cell death 1(DAD1) is a negative regulator of apoptosis and a subunit of the oligosaccharyltransferase complex.We cloned DAD1 from the spleen of Epinephelus awoara using the combinative methods of suppressive subtraction hybridization,SMART cDNA sy nthesis and RACE-PCR.The full-length cDNA of DAD1 is 721 bp,containing a 342 bp open reading frame that encodes a 113 ami no acid protein.Semi-quantitative RT-PCR analyses showed that mRNA of DAD1 can be detected in the spleen,heart,liver,anterior kidney and kidney of normal E.awoara and lipopolysaccharide-stimulated E.awoara.The expressions of DAD1 in all these tissues showed some increase in the LPS-treated E.awoara,compared with normal E.awoara,which indicates that lipopolysaccharide might be a stimulator of programmed cell death.
- defender against cell death 1 /
- cloning /
- expression /
- Epinephelus awoara

HTML全文

参考文献(25)

RAFF M C.Social control on cell survival and cell death[J].Nature,1992.356:397-400.

RAFF M.Cell suicide for beginners[J].Nature,1998,(396):119-122.

WILLIAMS G T,SMITH C A.Molecular regulation of apoptosis:genetic controls on cell death[J].Cell,1993,(7):777-779.

SCHWARTZAMA R A,CIDLOWSKI J A.Apoptosis:The biochemistry and molecular biology of programmed cell death[J].Endocr Rev,1993,(14):133 151.

STELLER H.Mechanisms and genes of cellular suicide[J].Science 1995(267):1 445-1 449.

VAUX D L,WEISSMAN I L,KIM S K.Prevention of programmed cell death in Caenorhabditis elegans by human bcl-2[J].Science,1992,(258):1 955-1 957.

NAKASHIMA T,SEKIGUCHI T,KURAOKA A,et al.Molecular cloning of a human cDNA encoding a novel protein,DAD1,whose defect causes apoptotic cell death in hamster BHK21 cells[J].Mol Cell Biol,1993,(13):6 367-6 374.

LEVINE A J.P53,the cellular gatekeeper for growth and division[J].Cell,1997,(88):323-331.

SILBERSTEINS P,COLLINS G,KELLEHER D J,et al.The essential OST2 gene encodes the 16-kDa subunit of the yeast oligosaccharyltransferase,a highly conserved protein expressed in diverse eukaryotic organisms[J].J Cell Biol,1995,131:371-383.

KELLEHER D,GILMORE R.DAD1,the defender against apoptotic cell death,is a subunit of the mammalian oligosaccharyltransferase[J].Proc Natl Acad Sci USA,1997,(94):4 994-4 999.

NISHII K,TSUZUKI T,KUMAI M,et al.Abnormalities of developmental cell death in Dad1-deficient mice[J].Genes Cells,1999,(4):243-252.

SUGIMOTO A,HOZAK R R,NAKASHIMA T,et al.DAD1,an endogenous programmed cell death suppressor in Caenorhabditis elegans and vertebrates[J].EMBO J,1995,(14):4 434-4 441.

APTE S S,MATTEI M G,SELDIN M F,et al.The highly conserved defender against the death (DAD1) gene maps to human chromosome 14q11-q12 and mouse chromosome 14 and has plant and nematode homologs[J].FEBS Lett,1995,(363):304-306.

DONG Y H,ZHAN X C,KVARNHEDEN A K,et al.Expression of a cDNA from apple encoding a homologue of DAD1,an inhibitor of programmed cell death[J].Plant Science,1998,(139):165-174.

GALLOIS P,MAKISHIMA T,HECHTT V,et al.An Arabidopsis thaliana cDNA complementing a hamster apoptosis suppressor mutant[J].J Plant,1997,(11):1 325-1 331.

THOMPSON J D,HIGGINS D G,GIBSON T J.Clustal W:improving the sensitivity of progressive multiple sequence alignment through sequence weighting,position-specific gap penalties and weight matrix choice.Nucleic Acids Res,1994;22:4 673-4 680.

SAITOU N,NEI M.The neighbor-joining method:a new method for reconstructing phylogenetic trees.Mol Biol Evol,1987,4:406-425.

MAKISHIMA T,NAKASHIMA T,NAGATA K K,et al.The highly conserved DAD1 protein involved in apoptosis is required for N-linked glycosylation[J].Genes Cells,1997,2:129.

BREWSTER J L,MARTIN S L,TOMS J,et al.Deletion of Dad1 in mice induces an apoptosis-associated embryonic death[J].Genesis,2000,(4):271 -278.

HONG N A,FLANNERY M,HSIEH S N,et al.Mice lacking Dad1,the defender against apoptotic death-1,express abnormal N-linked glycoproteins and undergo increased embryonic apoptosis[J].Dev Biol,2000,(1):76-84.

张成岗,贺福初.生物信息学方法与实践[M].北京:科学出版社,2002.63-93.

HONG N A,CADO D,MITCHELL J,et al.A targeted mutation at the T-cell receptor a/d locus impairs T-cell development and reveals the presence of the nearby antiapoptosis gene Dad1[J].Mol Cell Biol,1997,(17):2151.

LEE K S,CHUNG E H,HAN J H,et al.cDNA cloning of a defender against apoptotic cell death 1 (DAD1) homologue,responsive to external temperature stimulus from the spider,Araneus ventricosus[J].Comp Biochem Physiol B Biochem Mol Biol,2003,(1):117-123.

HONG N A,KABRA N H,HSIEH S N,et al.In vivo overexpression of Dad1,the defender against apoptotic death-1,enhances T cell proliferation but does not protect against apoptosis[J].J Immunol,1999,163:1 888.

TANAKA K,KONDOH N,SHUDA M,et al.Enhanced expression of mRNAs of antisecretory factor-1,gp96,DAD1 and CDC34 in human hepatocellular carcinomas[J].Biochim Biophys Acta,2001,(1):1-12.

施引文献

资源附件(0)

访问统计