Gong Li, Unnikrishnan Indira, Raghavan Anuradha, Parmar Kalindi, Rosenberg Naomi
Department of Molecular Biology and Microbiology, School of Medicine, Tufts University, Boston, Massachusetts 02111, USA.
J Virol. 2004 Feb;78(4):1636-44. doi: 10.1128/jvi.78.4.1636-1644.2004.
Suppression of apoptosis is an important feature of the Abelson murine leukemia virus (Ab-MLV) transformation process. During multistep transformation, Ab-MLV-infected pre-B cells undergo p53-dependent apoptosis during the crisis phase of transformation. Even once cells are fully transformed, an active v-Abl protein tyrosine kinase is required to suppress apoptosis because cells transformed by temperature-sensitive (ts) kinase mutants undergo rapid apoptosis after a shift to the nonpermissive temperature. However, inactivation of the v-Abl protein by a temperature shift interrupts signals transmitted via multiple pathways, making it difficult to identify those that are critically important for the suppression of apoptosis. To begin to dissect these pathways, we tested the ability of an SH2 domain Ab-MLV mutant, P120/R273K, to rescue aspects of the ts phenotype of pre-B cells transformed by the conditional kinase domain mutant. The P120/R273K mutant suppressed apoptosis at the nonpermissive temperature, a phenotype correlated with its ability to activate Akt. Apoptosis also was suppressed at the nonpermissive temperature by constitutively active Akt and in p53-null pre-B cells transformed with the ts kinase domain mutant. These data indicate that an intact Src homology 2 (SH2) domain is not critical for apoptosis suppression and suggest that signals transmitted through Akt and p53 play an important role in the response.
凋亡抑制是阿贝尔逊鼠白血病病毒(Ab-MLV)转化过程的一个重要特征。在多步骤转化过程中,Ab-MLV感染的前B细胞在转化的危机阶段经历p53依赖性凋亡。即使细胞完全转化后,仍需要有活性的v-Abl蛋白酪氨酸激酶来抑制凋亡,因为由温度敏感(ts)激酶突变体转化的细胞在转移到非允许温度后会迅速凋亡。然而,通过温度变化使v-Abl蛋白失活会中断通过多种途径传递的信号,从而难以确定那些对凋亡抑制至关重要的信号。为了开始剖析这些途径,我们测试了一种SH2结构域Ab-MLV突变体P120/R273K拯救由条件性激酶结构域突变体转化的前B细胞ts表型某些方面的能力。P120/R273K突变体在非允许温度下抑制凋亡,该表型与其激活Akt的能力相关。在非允许温度下,组成型活性Akt以及用ts激酶结构域突变体转化的p53缺失前B细胞也能抑制凋亡。这些数据表明完整的Src同源2(SH2)结构域对凋亡抑制并不关键,并提示通过Akt和p-53传递的信号在该反应中起重要作用。