Apoptotic DNA Fragmentation May Be a Cooperative Activity between Caspase-activated Deoxyribonuclease and the Poly(ADP-ribose) Polymerase-regulated DNAS1L3, an Endoplasmic Reticulum-localized Endonuclease That Translocates to the Nucleus during Apoptosis

被引：68

作者：

Errami, Youssef ^{[1
,2
]}

Naura, Amarjit S. ^{[1
,3
]}

Kim, Hogyoung ^{[1
]}

Ju, Jihang ^{[1
]}

Suzuki, Yasuhiro ^{[2
]}

El-Bahrawy, Ali H. ^{[1
,4
]}

Ghonim, Mohamed A. ^{[1
,5
]}

Hemeida, Ramadan A. ^{[4
]}

Mansy, Moselhy S. ^{[5
]}

Zhang, Jianhua ^{[6
]}

Xu, Ming ^{[7
]}

Smulson, Mark E. ^{[8
]}

Brim, Hassan ^{[9
,10
]}

Boulares, A. Hamid ^{[1
,2
]}

机构：

[1] Louisiana State Univ, Hlth Sci Ctr, Stanley Scott Canc Ctr, New Orleans, LA 70112 USA

[2] Louisiana State Univ, Hlth Sci Ctr, Dept Pharmacol & Expt Therapeut, New Orleans, LA 70112 USA

[3] Louisiana State Univ, Hlth Sci Ctr, Dept Med, New Orleans, LA 70112 USA

[4] Al Azhar Univ, Dept Pharmacol & Toxicol, Assiut 71515, Egypt

[5] Al Azhar Univ, Fac Pharm, Cairo 11787, Egypt

[6] Univ Alabama Birmingham, Dept Pathol, Birmingham, AL 35294 USA

[7] Univ Chicago, Dept Anesthesia & Crit Care, Chicago, IL 60637 USA

[8] Georgetown Univ, Sch Med, Dept Biochem, Washington, DC 20057 USA

[9] Howard Univ, Dept Pathol, Washington, DC 20059 USA

[10] Howard Univ, Ctr Canc, Washington, DC 20059 USA

来源：

JOURNAL OF BIOLOGICAL CHEMISTRY | 2013年 / 288卷 / 05期

基金：

美国国家卫生研究院;

关键词：

CHROMATIN CONDENSATION; CELLS; ROLES; THYMOCYTES; CLEAVAGE; IDENTIFICATION; DFF40/CAD; PROTEASES; DNASE1L3; CA2+;

D O I：

10.1074/jbc.M112.423061

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

070307 [化学生物学]; 071010 [生物化学与分子生物学];

摘要：

Caspase-activated DNase (CAD) is the most favorable candidate for chromatin degradation during apoptosis. Ca2+-dependent endonucleases are equally important in internucleosomal DNA fragmentation (INDF), including the PARP-1-regulated DNAS1L3. Despite the elaborate work on these endonucleases, the question of whether these enzymes cooperate during INDF was not addressed. Here, we show a lack of correlation between INDF and CAD expression levels and inactivation by cleavage of its inhibitor (ICAD) during apoptosis. The cells that failed to induce INDF accumulated large amounts of 50-kb breaks, which is suggestive of incomplete chromatin processing. Similarly, INDF was blocked by Ca2+ chelation without a block in ICAD cleavage or caspase-3 activation, which is consistent with the involvement of CAD in 50-kb DNA fragmentation and its Ca2+ independence. However, DNAS1L3 expression in INDF-deficient cells promoted INDF during apoptosis and was blocked by Ca2+ chelation. Interestingly, expression of DNAS1L3 in ICAD-deficient cells failed to promote tumor necrosis factor alpha-induced INDF but required the coexpression of ICAD. These results suggest a cooperative activity between CAD and DNAS1L3 to accomplish INDF. In HT-29 cells, endogenous DNAS1L3 localized to the endoplasmic reticulum (ER) and translocated to the nucleus upon apoptosis induction but prior to INDF manifestation, making it the first reported Ca2+-dependent endonuclease to migrate from the ER to the nucleus. The nuclear accumulation of DNAS1L3, but not its exit out of the ER, required the activity of cysteine and serine proteases. Interestingly, the endonuclease accumulated in the cytosol upon inhibition of serine, but not cysteine, proteases. These results exemplify the complexity of chromatin degradation during apoptosis.

引用

页码：3460 / 3468

页数：9

共 35 条

[1]

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[2]

Boulares AH, 2002, CANCER RES, V62, P4439

[3]

Roles of DNA fragmentation factor and poly(ADP-ribose) polymerase-1 in sensitization of fibroblasts to tumor necrosis factor-induced apoptosis [J].