Eskndir Nebiyat, Hossain Manseeb, Currey Marilena L, Pho Mai, Berrada Yasmin, Lin Katie, Manning Gianna, Prince Kelsey, Stephens Andrew D
Biology Department, University of Massachusetts Amherst, Amherst, MA 01003.
Molecular and Cellular Biology, University of Massachusetts Amherst, Amherst, MA 01003.
Mol Biol Cell. 2025 Mar 1;36(3):br6. doi: 10.1091/mbc.E24-05-0232. Epub 2024 Dec 20.
The nucleus must maintain stiffness to preserve its shape and integrity to ensure proper function. Defects in nuclear stiffness caused from chromatin and lamin perturbations produce abnormal nuclear shapes common in aging, heart disease, and cancer. Loss of nuclear shape via protrusions called blebs lead to nuclear rupture that is well established to cause nuclear dysfunction, including DNA damage. However, it remains unknown how increased DNA damage affects nuclear stiffness, shape, and ruptures, which could create a feedback loop. To determine whether increased DNA damage alters nuclear physical properties, we treated mouse embryonic fibroblast cells with DNA damage drugs cisplatin and bleomycin. DNA damage drugs caused increased nuclear blebbing and rupture in interphase nuclei within a few hours and independent of mitosis. Micromanipulation force measurements reveal that DNA damage decreased chromatin-based nuclear mechanics but did not change lamin-based strain stiffening at long extensions relative to wild type. Immunofluorescence measurements of DNA damage treatments reveal the mechanism is an ATM-dependent decrease in heterochromatin leading to nuclear weaken, blebbing, and rupture which can be rescued upon ATM inhibition treatment. Thus, DNA damage drugs cause ATM-dependent heterochromatin loss resulting in nuclear softening, blebbing, and rupture.
细胞核必须保持一定的硬度以维持其形状和完整性,从而确保正常功能。由染色质和核纤层扰动引起的核硬度缺陷会导致衰老、心脏病和癌症中常见的异常核形状。通过称为“泡”的突出物导致核形状丧失会引发核破裂,而核破裂会导致核功能障碍,包括DNA损伤,这一点已得到充分证实。然而,DNA损伤增加如何影响核硬度、形状和破裂尚不清楚,这可能会形成一个反馈回路。为了确定DNA损伤增加是否会改变核的物理性质,我们用DNA损伤药物顺铂和博来霉素处理小鼠胚胎成纤维细胞。DNA损伤药物在数小时内导致间期细胞核中核泡化和破裂增加,且与有丝分裂无关。显微操作力测量显示,相对于野生型,DNA损伤降低了基于染色质的核力学,但在长延伸时并未改变基于核纤层的应变硬化。DNA损伤处理的免疫荧光测量表明,其机制是ATM依赖性的异染色质减少,导致核弱化、泡化和破裂,而ATM抑制处理可挽救这种情况。因此,DNA损伤药物会导致ATM依赖性的异染色质丢失,从而导致核软化、泡化和破裂。