Patyal Pankaj, Ameer Fathima S, Verma Ambika, Zhang Xiaomin, Azhar Gohar, Shrivastava Jyotsna, Sharma Shakshi, Zhang Rachel, Wei Jeanne Y
Donald W. Reynolds Department of Geriatrics and Institute on Aging, University of Arkansas for Medical Sciences, Little Rock, AR 72205, USA.
Curr Issues Mol Biol. 2024 Aug 14;46(8):8835-8851. doi: 10.3390/cimb46080522.
The sirtuin-1 (SIRT1) gene contains multiple exons that usually undergo alternative splicing. The exclusion of one or more exons causes domain loss in the alternatively spliced isoforms and may change their functions. However, it is not completely established to what extent the loss of a non-catalytic domain could affect its regulatory function. Using muscle cells and SIRT1-knockout cells, we examined the function of the constitutively spliced isoform (SIRT1-v1) versus the alternatively spliced isoforms SIRT1-v2 and SIRT1-v3 that had lost part of the N-terminal region. Our data indicate that partial loss of the N-terminal domains in SIRT1-v2 and SIRT1-v3 attenuated their function. The full-length SIRT1-v1 significantly increased the oxidative phosphorylation and ATP production rate. Furthermore, SIRT1-v1 specifically upregulated the mitochondrial respiratory complex I without affecting the activity of complexes II, III, and IV. Additionally, domain loss affected the regulation of site-specific lysine acetylation in the histone H4 protein, the gene expression of respiratory complex I subunits, and the metabolic balance of oxidative phosphorylation versus glycolysis. Since alternatively spliced isoforms tend to increase with advancing age, the impact of SIRT1 isoforms on mitochondrial respiratory complexes warrants further investigation.
沉默调节蛋白1(SIRT1)基因包含多个外显子,这些外显子通常会发生可变剪接。一个或多个外显子的缺失会导致可变剪接异构体中的结构域丢失,并可能改变它们的功能。然而,非催化结构域的缺失在多大程度上会影响其调节功能尚未完全明确。我们使用肌肉细胞和SIRT1基因敲除细胞,研究了组成型剪接异构体(SIRT1-v1)与缺失部分N端区域的可变剪接异构体SIRT1-v2和SIRT1-v3的功能。我们的数据表明,SIRT1-v2和SIRT1-v3中N端结构域的部分缺失减弱了它们的功能。全长的SIRT1-v1显著提高了氧化磷酸化和ATP生成速率。此外,SIRT1-v1特异性地上调了线粒体呼吸复合体I,而不影响复合体II、III和IV的活性。此外,结构域的缺失影响了组蛋白H4蛋白中位点特异性赖氨酸乙酰化的调节、呼吸复合体I亚基的基因表达以及氧化磷酸化与糖酵解的代谢平衡。由于可变剪接异构体往往会随着年龄的增长而增加,SIRT1异构体对线粒体呼吸复合体的影响值得进一步研究。