Cytoskeleton integrity influences XRCC1 and PCNA dynamics at DNA damage

Abstract

Upon induction of DNA damage with 405 nm laser light, proteins involved in Base Excision Repair (BER) are recruited to DNA lesions. We find that the dynamics of factors typical of either short-patch (XRCC1) or long-patch (PCNA) BER are altered by chemicals that perturb actin or tubulin polymerization in human cells. Whereas the destabilization of actin filaments by Latrunculin B, Cytochalasin B or Jasplakinolide decreases BER factor accumulation at laser-induced damage, inhibition of tubulin polymerization by Nocodazole increases it. We detect no recruitment of actin to sites of laser-induced DNA damage, yet the depolymerization of cytoplasmic actin filaments elevates both actin and tubulin signals in the nucleus. While published evidence suggested a positive role for F-actin in double-strand break repair in mammals, the enrichment of actin in budding yeast nuclei interferes with BER, augmenting sensitivity to Zeocin. Our quantitative imaging results suggest that the depolymerization of cytoplasmic actin may compromise BER efficiency in mammals not only due to elevated levels of nuclear actin, but also of tubulin. Our study is one of few linking cytoskeletal integrity to BER. [Media: see text] [Media: see text]

Persistent confined migration confers permanent nuclear and functional changes in migrating cells

“…To further confirm this point, we performed in situ DNA digestion and observed that control cells were more sensitive to DNA digestion (visualized as a reduction in the nuclear area by DNAse activity) than MA cells (Figure S6G). Finally, several reports show that actin cytoskeleton is required during DNA damage response [36,37]. We investigated the levels of γH2AX in MA cells treated with latrunculin and jasplakinolide, and found that γH2AX were reduced in MA cells treated with latrunculin, whereas jasplakinolide treatment exhibited a more similar phenotype to untreated MA cells (Figures 6F, and 6G).…”

DNA Damage Response Gene-Based Subtypes Associated With Clinical Outcomes in Early-Stage Lung Adenocarcinoma

“…Many studies have reported that they are significantly correlated with prognosis in different cancer types, such as colorectal cancer, pancreatic cancer, gastric cancer, and so on ( Luo et al, 2018 ; Zhang et al, 2019 ; Li et al, 2021 ). NEIL3 , PCNA , RFC3 , and POLD2 play important roles in the BER pathway which are recruited to DNA lesions and cleave and repair the damaged bases cooperatively ( Robertson et al, 2009 ; Hurst et al, 2021 ; Wang et al, 2021 ). Zhao et al found that NEIL3 activated cell cycle progression, leading to poor prognosis ( Zhao et al, 2021 ).…”

Molecular mechanism behind methamphetamine‐induced damages in testicular tissue: Evidences for oxidative stress, autophagy, and apoptosis

“…Indeed, at first stage of ODD, the PCNA in cooperation with BER enzymes (mainly glycosylases), initiates the repairing process. Thus, failure of this system ends up with expanded DNA damage in the testicular cellularity (Hurst et al, 2021; Moradi‐Ozarlou et al, 2021; Moshari et al, 2017; Tsai et al, 2017). Therefore, the METH‐suppressed PCNA expression might negatively affect BER enzyme expression.…”

“…Indeed, proliferating cell nuclear antigen (PCNA) is a member of the sliding clamp family, which involves as a proceeding element in the polymerase process (Moradi-Ozarlou et al, 2021;Moshari et al, 2017) and is reported to promote the BER enzymes-induced roles in the DNA repairing process. In line with this issue, it has shown that the PCNA plays a crucial role in interacting with almost all players in the BER process, including glycosylases, AP endonuclease, and XRCC1 (Hurst et al, 2021;Tsai et al, 2017). As a matter of fact, the BER pathway removes the DNA lesions mainly after ODD, which is divided into different stages of detection and cutting, incision, replacement, processing of DNA terminals, and DNA sealing (Hu et al, 2021).…”