Smith Chad, D'Mello Santosh R
Department of Biological Sciences, Southern Methodist University, Dedman Life Sciences Building, 6501 Airline Road, Dallas, TX, 75275, USA.
Department of Molecular and Cell Biology, University of Texas at Dallas, Richardson, TX, 75080, USA.
Mol Neurobiol. 2016 Oct;53(8):5628-39. doi: 10.1007/s12035-015-9452-3. Epub 2015 Oct 17.
Previous studies performed in cell lines have shown that the heat shock protein, DNAJB6, protects against the proteotoxic effects of mutant huntingtin (mut-Htt) via direct interaction with mut-Htt. However, these studies were performed primarily using in vitro models and cell lines. We report that when expressed in primary neurons, DNAJB6 induces cell death. Neurotoxicity is observed with both the DNAJB6a isoform, which is strictly nuclear, and the DNAJB6b isoform, which is predominantly cytoplasmic, suggesting that neurotoxicity is mediated in the nucleus. However, when co-expressed in primary neurons with mut-Htt, DNAJB6 protects against mut-Htt neurotoxicity. This suggests that the contrasting effect of DNAJB6 on neuronal viability depends on the presence or absence of proteotoxic stress. Neurotoxicity of DNAJB6 cannot be prevented by inhibition of glycogen synthase kinase 3 beta (GSK3β) or c-Jun N-terminal kinase (JNK) but is prevented by pharmacological inhibition of cyclin-dependent kinases (CDKs). Expression of dominant-negative forms of CDK2 or CDK4, or of p21(CIP1), the physiological inhibitor of CDKs, also inhibits DNAJB6 neurotoxicity. DNAJB6 neurotoxicity can also be inhibited by histone deacetylase-4 (HDAC4), which interacts with DNAJB6 and which has previously been described to inhibit cell cycle progression. These results conclude that neurotoxicity resulting from elevated DNAJB6 is cell cycle dependent.
先前在细胞系中进行的研究表明,热休克蛋白DNAJB6通过与突变型亨廷顿蛋白(mut-Htt)直接相互作用,保护细胞免受mut-Htt的蛋白毒性作用。然而,这些研究主要是使用体外模型和细胞系进行的。我们报告称,当在原代神经元中表达时,DNAJB6会诱导细胞死亡。无论是严格定位于细胞核的DNAJB6a亚型,还是主要位于细胞质的DNAJB6b亚型,均会观察到神经毒性,这表明神经毒性是在细胞核中介导的。然而,当在原代神经元中与mut-Htt共表达时,DNAJB6可保护细胞免受mut-Htt的神经毒性。这表明DNAJB6对神经元活力的相反作用取决于蛋白毒性应激的存在与否。抑制糖原合酶激酶3β(GSK3β)或c-Jun氨基末端激酶(JNK)并不能预防DNAJB6的神经毒性,但细胞周期蛋白依赖性激酶(CDK)的药理学抑制可预防这种毒性。CDK2或CDK4的显性负性形式,或CDK的生理抑制剂p21(CIP1)的表达,也可抑制DNAJB6的神经毒性。DNAJB6的神经毒性还可被组蛋白脱乙酰基酶-4(HDAC4)抑制,HDAC4与DNAJB6相互作用,此前已被描述为可抑制细胞周期进程。这些结果表明,DNAJB6水平升高导致的神经毒性是细胞周期依赖性的。