Gregory M A, Hann S R
Department of Cell Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-2175, USA.
Mol Cell Biol. 2000 Apr;20(7):2423-35. doi: 10.1128/MCB.20.7.2423-2435.2000.
The c-Myc oncoprotein is a transcription factor which is a critical regulator of cellular proliferation. Deregulated expression of c-Myc is associated with many human cancers, including Burkitt's lymphoma. The c-Myc protein is normally degraded very rapidly with a half-life of 20 to 30 min. Here we demonstrate that proteolysis of c-Myc in vivo is mediated by the ubiquitin-proteasome pathway. Inhibition of proteasome activity blocks c-Myc degradation, and c-Myc is a substrate for ubiquitination in vivo. Furthermore, an increase in c-Myc stability occurs in mitotic cells and is associated with inhibited c-Myc ubiquitination. Deletion analysis was used to identify regions of the c-Myc protein which are required for rapid proteolysis. We found that a centrally located PEST sequence, amino acids 226 to 270, is necessary for rapid c-Myc degradation, but not for ubiquitination. Also, N-terminal sequences, located within the first 158 amino acids of c-Myc, are necessary for both efficient c-Myc ubiquitination and subsequent degradation. We found that c-Myc is significantly stabilized (two- to sixfold) in many Burkitt's lymphoma-derived cell lines, suggesting that aberrant c-Myc proteolysis may play a role in the pathogenesis of Burkitt's lymphoma. Finally, mutation of Thr-58, a major phosphorylation site in c-Myc and a mutational hot spot in Burkitt's lymphoma, increases c-Myc stability; however, mutation of c-Myc is not essential for stabilization in Burkitt's lymphoma cells.
c-Myc癌蛋白是一种转录因子,是细胞增殖的关键调节因子。c-Myc的表达失调与许多人类癌症相关,包括伯基特淋巴瘤。c-Myc蛋白通常降解非常迅速,半衰期为20至30分钟。在此我们证明,c-Myc在体内的蛋白水解是由泛素-蛋白酶体途径介导的。蛋白酶体活性的抑制会阻止c-Myc的降解,并且c-Myc是体内泛素化的底物。此外,c-Myc的稳定性在有丝分裂细胞中增加,并且与c-Myc泛素化的抑制相关。缺失分析用于鉴定c-Myc蛋白中快速蛋白水解所需的区域。我们发现,位于中央的PEST序列(氨基酸226至270)对于c-Myc的快速降解是必需的,但对于泛素化不是必需的。同样,位于c-Myc前158个氨基酸内的N端序列对于有效的c-Myc泛素化和随后的降解都是必需的。我们发现,在许多伯基特淋巴瘤来源的细胞系中,c-Myc显著稳定(两到六倍),这表明异常的c-Myc蛋白水解可能在伯基特淋巴瘤的发病机制中起作用。最后,Thr-58(c-Myc中的主要磷酸化位点和伯基特淋巴瘤中的突变热点)的突变增加了c-Myc的稳定性;然而,c-Myc的突变对于伯基特淋巴瘤细胞中的稳定化不是必需的。