dNTP Pool Imbalance Induced Endonuclease : Mechanism of All Death.
dNTP Pool Imbalance Induced Endonuclease : Mechanism of All Death.
批准号:
03807146
负责人:
WATAYA Yusuke
金额:
$1.15万
依托单位:
依托单位国家:
日本
项目类别:
Grant-in-Aid for General Scientific Research (C)
财政年份:
1991
资助国家:
日本
项目状态:
已结题
起止时间:
1991 至 1992
中文摘要
研究了细胞内脱氧核苷三磷酸(dNTP)库失衡诱导小鼠FM 3A细胞死亡的机制。5-氟脱氧尿苷(FdUrd)可诱导细胞内dNTP池失衡,导致DNA双链断裂和细胞死亡;放线菌酮(cycloheximide)可抑制FdUrd诱导的DNA双链断裂和细胞死亡;在这个实验中,我们已经从FdUrd处理的FM 3A细胞中纯化并表征了双链核酸内切酶。用柱层析法将酶纯化至接近均一。通过含有DNA的SDS-PAGE,核酸内切酶的分子量约为45 kDa。核酸内切酶不需要二价金属阳离子,并且随机切割裸双链体DNA。此外,我们还研究了FdUrd诱导的DNA片段化。FdUrd诱导的DNA片段分为两类。一个是100-200 kbp的大片段,另一个是180 bp左右的梯状DNA片段,其节距与核小体长度相当。在第二种情况下,DNA片段化被认为是细胞凋亡的特征类型。此外,我们发现,核原癌基因,c-fos,c-jun和c-myc的mRNA水平增加,在FdUrd处理的细胞。我们得出结论,纯化的核酸内切酶是一种新的核酸内切酶,不同于以前的特点哺乳动物核酸内切酶。这种酶被称为内切核酸酶K。
英文摘要
The mechanism of intracellular deoxyribonucleoside triphosphates (dNTP) pool imbalance-induced cell death in mouse FM3A cells was studied. We previously reported that 1) 5-fluoro-deoxyuridine (FdUrd) induced intracellular dNTP pool imbalance followed by DNA double strand breaks and subsequent cell death, 2) these DNA double strand breaks and cell death were inhibited by cycloheximide, an inhibitor of protein biosynthesis, 3) an endonuclease activity toward double stranded DNA was detected in the lysate ofFdUrd-treated FM3A cells but not untreated cells. In this experiment, we have purified and characterized the double-stranded endonuclease from FdUrd-treated FM3A cells. The enzyme which was purified to near homogeneity by column chromatographys. The molecular mass of the endonuclease was approximately 45 kDa by SDS-PAGE that contained DNA. The endonuclease required no divalent metal cations and cleaved naked duplex DNA at random. In addition we have investigated the DNA fragmentation induced by FdUrd. FdUrd-induced DNA fragments were separated into two classes. One was large DNA fragment with size of 100-200 kbp, and other was ladder DNA fragment with pitch of nucleosome length, approximately 180 bp. In second case the DNA fragmentation was of the type considered characteristic of apoptosis. Furthermore, we found that mRNA levels of nuclear proto-oncogenes, c-fos, c-jun and c-myc, increased in FdUrd-treated cells. We conclude that the purified endonuclease is a novel endonuclease, distinct from previously characterized mammalian endonucleases. The enzyme, which was called endonuclease K.
期刊论文(38)
专著(0)
科研奖励(0)
会议论文
登录
查看更多内容
M.Otani, S.Yoshida, A.Yoshioka-Hiramoto, T.Nakazawa, and Y.Wataya: "The dNTP Imbalance Death" Nucleic Acids Symposium Series. No.25. 111-112 (1991)
M.Otani、S.Yoshida、A.Yoshioka-Hiramoto、T.Nakazawa 和 Y.Wataya:“dNTP 不平衡死亡”核酸研讨会系列。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
M.Yamato.J.Ando,K.Sakata,K.Haohigaki,Y.Wataya,A.Tsukagoshi,T.Taslizo&T.Tsuruta: "Synthesis and Anyitumor Activity of Tro polone Derivations 7.Bistropolone Containing Connecting Methylene Chain" J.Med.Chem. 35. 267-273 (1992)
M.Yamato.J.Ando、K.Sakata、K.Haohigaki、Y.Wataya、A.Tsukagoshi、T.Taslizo
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
H.Satake: "Action of 5-Trifluoromethyl-2^2-deoxyuridine on DNA 5ynthesis." Nucleic Acids Symposium Series. 26. 193-194 (1992)
H.Satake:“5-三氟甲基-2^2-脱氧尿苷对 DNA 5 合成的作用。”
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
S.Yoshida, K.Watanabe, A.Yoshioka-Hiramoto, and Y.Wataya: "The dNTP Imbalance Death" Nucleisides and Nucleotides.
S.Yoshida、K.Watanabe、A.Yoshioka-Hiramoto 和 Y.Wataya:“dNTP 不平衡死亡”核苷和核苷酸。
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
M.Yamato,Y.Hirota,S.Yoshida,S.Tanaka,T.Morita,J.Sakai K.Hadigaki,H.Hagatsu and Y.Wataya: "Imbalomce of Desxpribomucleoside Triphotes and DNA Double StaanlーBraks in Mouse Mammary Tumor FM3Aalls Treatel in vitro with an Antineoplastic fropolone Derirative"
M. Yamato、Y. Hirota、S. Yoshida、S. Tanaka、T. Morita、J. Sakai K. Hadigaki、H. Hagatsu 和 Y. Wataya:“小鼠乳腺肿瘤 FM3Aalls 中 Desxpribomucleoside Triphotes 和 DNA Double Staanl Braks 的不平衡抗肿瘤福罗泊酮衍生物的体外治疗"
DOI:
--
发表时间:
期刊:
影响因子:
--
作者:
[]
通讯作者:
共 28 条
Study of novel anti-leishmanial drug
-
批准号:23659212
-
项目类别:Grant-in-Aid for Challenging Exploratory Research
-
资助金额:$2.33万
-
财政年份:2011
-
负责人:WATAYA Yusuke
-
依托单位:
Study of new broad spectrum of anti-parasitic agent and basis of molecular base of it
-
批准号:22390024
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$11.23万
-
财政年份:2010
-
负责人:WATAYA Yusuke
-
依托单位:
New antiamlarial drug development research including mechanism analysis of drug-resistant Plasmodium falciparum
-
批准号:14021072
-
项目类别:Grant-in-Aid for Scientific Research on Priority Areas
-
资助金额:$35.2万
-
财政年份:2002
-
负责人:WATAYA Yusuke
-
依托单位:
New Antimalarial Drug Research for Multidrug-resistant Malaria
-
批准号:12307007
-
项目类别:Grant-in-Aid for Scientific Research (A)
-
资助金额:$23.18万
-
财政年份:2000
-
负责人:WATAYA Yusuke
-
依托单位:
Development of simple and specific DNA diagnostic method of Malaria in Malaria endemic areas
-
批准号:11557183
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$8.06万
-
财政年份:1999
-
负责人:WATAYA Yusuke
-
依托单位:
Study of the molecular mechanisms of cell death induced by the dNTP pool imbalance.
-
批准号:08457607
-
项目类别:Grant-in-Aid for Scientific Research (B)
-
资助金额:$4.93万
-
财政年份:1996
-
负责人:WATAYA Yusuke
-
依托单位:
Development of Anti-malarial Drug and it's Molecular Target
-
批准号:08281105
-
项目类别:Grant-in-Aid for Scientific Research on Priority Areas (A)
-
资助金额:$124.16万
-
财政年份:1996
-
负责人:WATAYA Yusuke
-
依托单位:
Development of a new DNA diagnostic system for the species-specific detection of human malaria parasites using specific nucleotide sequences of the 18S ribosomal RNA gene.
-
批准号:07557301
-
项目类别:Grant-in-Aid for Scientific Research (A)
-
资助金额:$2.75万
-
财政年份:1995
-
负责人:WATAYA Yusuke
-
依托单位:
dNTP Imbalance and DNA Double Strand Breaks in Mouse FM3A Cells and the Mechanism of Cell Death
-
批准号:05807206
-
项目类别:Grant-in-Aid for General Scientific Research (C)
-
资助金额:$1.22万
-
财政年份:1993
-
负责人:WATAYA Yusuke
-
依托单位:
DNA Diagnosis of Malaria Using PCR Techniques.
-
批准号:03557020
-
项目类别:Grant-in-Aid for Developmental Scientific Research (B)
-
资助金额:$5.82万
-
财政年份:1991
-
负责人:WATAYA Yusuke
-
依托单位:
Deoxyribonucleoside Triphosphate Imbalance: The Mechanism of Cell Death and DNA-Double Strand Breaks.
-
批准号:62570989
-
项目类别:Grant-in-Aid for General Scientific Research (C)
-
资助金额:$1.34万
-
财政年份:1987
-
负责人:WATAYA Yusuke
-
依托单位:
Anti-Parasitic Activity of Nucleoside Analogues against Leishmania tropica and Leishmania donovani.
-
批准号:61870087
-
项目类别:Grant-in-Aid for Developmental Scientific Research
-
资助金额:$3.71万
-
财政年份:1986
-
负责人:WATAYA Yusuke
-
依托单位:
海外基金