Bloomfield Center for Research in Aging, Lady Davis Institute for Medical Research, Sir Mortimer B. Davis Jewish General Hospital, Montréal, Canada.
PLoS One. 2009 Aug 14;4(8):e6647. doi: 10.1371/journal.pone.0006647.
Previously, we have shown the loss of anti-Bax function in Creutzfeldt Jakob disease (CJD)-associated prion protein (PrP) mutants that are unable to generate cytosolic PrP (CyPrP). To determine if the anti-Bax function of PrP modulates the manifestation of prion diseases, we further investigated the anti-Bax function of eight familial Gerstmann-Sträussler-Scheinker Syndrome (GSS)-associated PrP mutants. These PrP mutants contained their respective methionine ((M)) or valine ((V)) at codon 129. All of the mutants lost their ability to prevent Bax-mediated chromatin condensation or DNA fragmentation in primary human neurons. In the breast carcinoma MCF-7 cells, the F198S(V), D202N(V), P102L(V) and Q217R(V) retained, whereas the P102L(M), P105L(V), Y145stop(M) and Q212P(M) PrP mutants lost their ability to inhibit Bax-mediated condensed chromatin. The inhibition of Bax-mediated condensed chromatin depended on the ability of the mutants to generate cytosolic PrP. However, except for the P102L(V), none of the mutants significantly inhibited Bax-mediated caspase activation. These results show that the cytosolic PrP generated from the GSS mutants is not as efficient as wild type PrP in inhibiting Bax-mediated cell death. Furthermore, these results indicate that the anti-Bax function is also disrupted in GSS-associated PrP mutants and is not associated with the difference between CJD and GSS.
先前,我们已经证明了无法生成细胞质朊蛋白 (CyPrP) 的 Creutzfeldt-Jakob 病 (CJD) 相关朊蛋白 (PrP) 突变体中抗 Bax 功能的丧失。为了确定 PrP 的抗 Bax 功能是否调节朊病毒病的表现,我们进一步研究了 8 种家族性格斯特曼-施特劳斯勒-谢因克综合征 (GSS) 相关 PrP 突变体的抗 Bax 功能。这些 PrP 突变体在密码子 129 处含有各自的蛋氨酸 ((M)) 或缬氨酸 ((V))。所有突变体均丧失了防止 Bax 介导的人原代神经元染色质浓缩或 DNA 片段化的能力。在乳腺癌 MCF-7 细胞中,F198S(V)、D202N(V)、P102L(V)和 Q217R(V)保留了抗 Bax 介导的浓缩染色质的能力,而 P102L(M)、P105L(V)、Y145stop(M)和 Q212P(M)PrP 突变体丧失了抑制 Bax 介导的浓缩染色质的能力。抗 Bax 介导的浓缩染色质的抑制取决于突变体产生细胞质 PrP 的能力。然而,除了 P102L(V) 之外,没有一个突变体显著抑制 Bax 介导的半胱天冬酶激活。这些结果表明,从 GSS 突变体产生的细胞质 PrP 在抑制 Bax 介导的细胞死亡方面不如野生型 PrP 有效。此外,这些结果表明抗 Bax 功能也在 GSS 相关 PrP 突变体中被破坏,并且与 CJD 和 GSS 之间的差异无关。