Mechanism of Apoptosis Induction by Mycoplasmal Nuclease MGA_0676 in Chicken Embryo Fibroblasts.
Mechanism of Apoptosis Induction by Mycoplasmal Nuclease MGA_0676 in Chicken Embryo Fibroblasts.
复制标题
支原体核酸酶MGA_0676诱导鸡胚成纤维细胞凋亡的机制
DOI：
10.3389/fcimb.2018.00105
复制
发表时间：
2018
影响因子：
5.7
通讯作者：
Wu WX
中科院分区：
文献类型：
--
作者：
Li P;Xu J;Rao HM;Li X;Zhang YK;Jiang F;Wu WX
MGA_0676已被表征为可诱导鸡细胞凋亡的鸡毒支原体核酸酶。然而，MGA_0676诱导细胞凋亡的机制仍不清楚。在这项研究中，我们评估了MGA_0676诱导的永生化鸡胚成纤维细胞（DF-1）和癌细胞系的凋亡和内化。通过小窝蛋白介导的内吞作用，通过用特异性抑制剂或siRNA阻断内吞作用，证实了MGA_0676的内化。我们鉴定了DF-1中NEDD 8激活酶E1调节亚基（NAE）的Thif结构域为与MGA_0676的SNC结构域相互作用的靶区域。观察到Thif和SNC结构域之间的相互作用共同位于DF-1的核周和核中。我们发现NAE和MGA_0676的相互作用增强了DF-1细胞的凋亡能力，加速了cullin neddylation的过程，进而激活了NF-κB。这导致观察到NF-κB在DF-1细胞核中聚集。当siRNA或BAY 11-7082抑制NF-κB或siRNA沉默NAE时，MGA 0676诱导的细胞凋亡明显减少。总之，我们的研究结果表明，MGA_0676通过小窝蛋白介导的内吞作用内化，与SNC依赖性Thif相互作用以加速cullin neddylation过程并激活DF-1细胞中的NF-κB，最终在鸡细胞凋亡中发挥关键作用。我们的研究结果表明，MGA_0676构成了一个关键的致病性毒力因子的呼吸道疾病引起的M。鸡败血症这项研究还开辟了一个研究MGA_0676作为癌症研究中促凋亡药物的潜在候选者的场所。
MGA_0676 has been characterized as a Mycoplasma gallisepticum nuclease that can induce apoptosis of chicken cells. However, the mechanism by which MGA_0676 induces apoptosis has remained unclear. In this study, we evaluated MGA_0676-induced apoptosis and internalization in immortalized chicken embryo fibroblasts (DF-1) and cancer cell lines. The internalization of MGA_0676 was proven through caveolin-mediated endocytosis by blocking the endocytosis with specific inhibitors or with siRNA. We identified the Thif domain of NEDD8-activating enzyme E1 regulatory subunit (NAE) in DF-1 as the target region interacting with the SNC domain of MGA_0676. The interaction between the Thif and SNC domains was observed co-located in the perinuclear and nuclear of DF-1. We found that the interaction between NAE and MGA_0676 increased the ability of apoptosis and accelerated the process of cullin neddylation in DF-1 cells, in turn activating NF-κB. This resulted in the observed aggregation of NF-κB in the nuclei of DF-1 cells. Moreover, the apoptosis induced by MGA_0676 decreased significantly when NF-κB was inhibited by siRNA or BAY 11-7082 or when NAE was silenced by siRNA. Overall, our results demonstrate that MGA_0676 is internalized through caveolin-mediated endocytosis, interacts with SNC-dependent Thif to accelerate the process of cullin neddylation and activates NF-κB in DF-1 cells, ultimately playing a key role in apoptosis in chicken cells. Our results indicate MGA_0676 constitutes a critical etiological virulence factor of the respiratory disease caused by M. gallisepticum. This study also opens a venue to investigate MGA_0676 as a potential candidate as pro-apoptotic drug in cancer studies.
登录
查看更多内容
影响因子：
5.2
作者：
Krachler AM;Orth K
通讯作者：
Orth K
影响因子：
9.8
作者：
Chaudhary N;Gomez GA;Howes MT;Lo HP;McMahon KA;Rae JA;Schieber NL;Hill MM;Gaus K;Yap AS;Parton RG
通讯作者：
Parton RG
影响因子：
6.7
作者：
Jubelin G;Taieb F;Duda DM;Hsu Y;Samba-Louaka A;Nobe R;Penary M;Watrin C;Nougayrède JP;Schulman BA;Stebbins CE;Oswald E
通讯作者：
Oswald E
影响因子：
2.6
作者：
Fu P;Sun Z;Zhang Y;Yu Z;Zhang H;Su D;Jiang F;Wu W
通讯作者：
Wu W
影响因子：
4.8
作者：
Chen, YZ;McPhie, DL;Neve, RL
通讯作者：
Neve, RL