Transient expression of the Bcl-2 family member, A1-a, results in nuclear localization and resistance to staurosporine-induced apoptosis.
Transient expression of the Bcl-2 family member, A1-a, results in nuclear localization and resistance to staurosporine-induced apoptosis.
复制标题
Bcl-2 家族成员 A1-a 的瞬时表达导致核定位和对十字孢菌素诱导的细胞凋亡的抵抗。
DOI：
10.1038/sj.cdd.4400879
复制
发表时间：
2001
期刊：
影响因子：
--
通讯作者：
Prystowsky,MB
中科院分区：
文献类型：
--
作者：
Somogyi,RD;Wu,Y;Orlofsky,A;Prystowsky,MB
Bcl2家族蛋白具有抗凋亡或促凋亡活性。通过确定Bcl2家族成员在细胞内的定位，已经深入了解了它们的功能。我们已经制备了抗A1-a的单抗，并使用COS-7过表达系统研究了小鼠抗凋亡的Bcl-2家族成员A1-a的定位。亚细胞分离和免疫荧光显微镜检测发现，A1-a在定位于细胞核的COS-7细胞中高表达。COS-7核中的A1-a与核基质紧密结合，表现为对DNase I和RNaseA处理以及1.0%Triton X-100、0.15M NaC l、0.25M HCl、0.5M Tris pH 7.4和6M尿素连续提取的抵抗。在十二烷基硫酸钠提取后，对A1-a进行高效液相分析，得到的组分在Western印迹分析中显示出多条带，这表明有形成多聚体的倾向。转导A1-a基因的COS-7细胞不受星形孢子素诱导的细胞凋亡的影响。细胞死亡与分化(2001)8,785-793
The Bcl-2 family of proteins has been characterized by either anti-apoptotic or pro-apoptotic activity. Insight into how Bcl-2 family members function has been gained by determining their intracellular localization. We have generated a monoclonal anti-A1-a antibody and used a COS-7 overexpression system to study the localization of the murine anti-apoptotic Bcl-2 family member, A1-a. A1-a overexpressed in COS-7 cells localized to the nucleus as determined by subcellular fractionation and immunofluorescent microscopy. A1-a in the COS-7 nucleus bound tightly to the nuclear matrix as evidenced by resistance to treatment with DNAse I and RNAse A and sequential extraction with 1.0% Triton X-100, 0.15 M NaCl, 0.25 M HCl, 0.5 M Tris pH 7.4 and 6 M urea. HPLC analysis of A1-a, subsequent to SDS extraction, produced fractions that gave multiple bands when analyzed by Western blot analysis suggesting a propensity to form multimers. COS-7 cells transfected with A1-a were protected from apoptotic induction by staurosporine treatment. Cell Death and Differentiation (2001) 8, 785–793