Joudeh Liza A, Logan Schuck P, Van Nina M, DiCintio Alannah J, Stewart Jason A, Waldman Alan S
Department of Biological Sciences, University of South Carolina, Columbia, SC 20208.
Department of Biology, Western Kentucky University, Bowling Green, KY 42101.
bioRxiv. 2024 Jul 2:2024.07.02.601729. doi: 10.1101/2024.07.02.601729.
Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare genetic condition characterized by features of accelerated aging, and individuals with HGPS seldom live beyond their mid-teens. The syndrome is commonly caused by a point mutation in the LMNA gene which codes for lamin A and its splice variant lamin C, components of the nuclear lamina. The mutation causing HGPS leads to production of a truncated, farnesylated form of lamin A referred to as "progerin." Progerin is also expressed at low levels in healthy individuals and appears to play a role in normal aging. HGPS is associated with an accumulation of genomic DNA double-strand breaks (DSBs) and alterations in the nature of DSB repair. The source of DSBs in HGPS is often attributed to stalling and subsequent collapse of replication forks in conjunction with faulty recruitment of repair factors to damage sites. In this work, we used a model system involving immortalized human cell lines to investigate progerin-induced genomic damage. Using an immunofluorescence approach to visualize phosphorylated histone H2AX foci which mark sites of genomic damage, we report that cells engineered to express progerin displayed a significant elevation of endogenous damage in the absence of any change in the cell cycle profile or doubling time of cells. Genomic damage was enhanced and persistent in progerin-expressing cells treated with hydroxyurea. Overexpression of wild-type lamin A did not elicit the outcomes associated with progerin expression. Our results show that DNA damage caused by progerin can occur independently from global changes in replication or cell proliferation.
哈钦森-吉尔福德早衰综合征(HGPS)是一种罕见的遗传疾病,其特征为加速衰老,患有HGPS的个体很少能活过青少年中期。该综合征通常由LMNA基因中的一个点突变引起,该基因编码核纤层蛋白A及其剪接变体核纤层蛋白C,它们是核纤层的组成部分。导致HGPS的突变会产生一种截短的、法尼基化形式的核纤层蛋白A,称为“早老素”。早老素在健康个体中也有低水平表达,并且似乎在正常衰老过程中发挥作用。HGPS与基因组DNA双链断裂(DSB)的积累以及DSB修复性质的改变有关。HGPS中DSB的来源通常归因于复制叉的停滞和随后的崩溃,以及修复因子错误募集到损伤位点。在这项工作中,我们使用了一个涉及永生化人类细胞系的模型系统来研究早老素诱导的基因组损伤。通过免疫荧光方法可视化标记基因组损伤位点的磷酸化组蛋白H2AX焦点,我们报告称,经基因工程改造以表达早老素的细胞在细胞周期分布或细胞倍增时间没有任何变化的情况下,内源性损伤显著升高。在用羟基脲处理的表达早老素的细胞中,基因组损伤增强且持续存在。野生型核纤层蛋白A的过表达并未引发与早老素表达相关的结果。我们的结果表明,早老素引起的DNA损伤可以独立于复制或细胞增殖的全局变化而发生。