β2-spectrin depletion impairs DNA damage repair

Horikoshi, Nobuo; Pandita, Raj K.; Mujoo, Kalpana; Hambarde, Shashank; Sharma, Dharmendra; Mattoo, Abid R.; Chakraborty, Sharmistha; Charaka, Vijaya; Hunt, Clayton R.; Pandita, Tej K.

doi:10.18632/oncotarget.9677

Cited by 19 publications

(32 citation statements)

References 64 publications

(90 reference statements)

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“…6C). To determine whether the decrease in RPA and Rad51 focus formation in MCL-1-depleted cells postirradiation was due to decreased ssDNA production as a result of defective resection, we did a quantitative resection assay using the ER-AsiSI system (23). In this resection assay system, MRE11-depleted cells were used as a positive control for loss of resection.…”

Section: Mcl-1 In Dna Damage Repair Molecular and Cellular Biologymentioning

confidence: 99%

MCL-1 Depletion Impairs DNA Double-Strand Break Repair and Reinitiation of Stalled DNA Replication Forks

Mattoo

Pandita

Chakraborty

et al. 2017

Molecular and Cellular Biology

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Myeloid cell leukemia 1 (MCL-1) is a prosurvival BCL-2 protein family member highly expressed in hematopoietic stem cells (HSCs) and regulated by growth factor signals that manifest antiapoptotic activity. Here we report that depletion of MCL-1 but not its isoform MCL-1S increases genomic instability and cell sensitivity to ionizing radiation (IR)-induced death. MCL-1 association with genomic DNA increased postirradiation, and the protein colocalized with 53BP1 foci. Postirradiation, MCL-1-depleted cells exhibited decreased ␥-H2AX foci, decreased phosphorylation of ATR, and higher levels of residual 53BP1 and RIF1 foci, suggesting that DNA double-strand break (DSB) repair by homologous recombination (HR) was compromised. Consistent with this model, MCL-1-depleted cells had a reduced frequency of IRinduced BRCA1, RPA, and Rad51 focus formation, decreased DNA end resection, and decreased HR repair in the DR-GFP DSB repair model. Similarly, after HU induction of stalled replication forks in MCL-1-depleted cells, there was a decreased ability to subsequently restart DNA synthesis, which is normally dependent upon HRmediated resolution of collapsed forks. Therefore, the present data support a model whereby MCL-1 depletion increases 53BP1 and RIF1 colocalization at DSBs, which inhibits BRCA1 recruitment, and sensitizes cells to DSBs from IR or stalled replication forks that require HR for repair.KEYWORDS MCL-1, BCL-2, HR, ICL, apoptosis, 53BP1, DSB repair M CL-1 is a member of the prosurvival BCL-2 family and plays an important role in the regulation of the intrinsic or mitochondrial apoptotic pathway by inhibiting both BH3-only proteins and the proapoptotic proteins (1-3). MCL-1 is mainly located at the outer mitochondrial membrane and inhibits the progression of apoptosis by sequestering executioner proapoptotic proteins BAK and BAX, which are capable of inducing pore formation in the mitochondrial membrane. The subsequent release of cytochrome c into the cytoplasm activates caspases which are responsible for the majority of the macromolecular degradation observed during apoptosis (3). Suppression of BAK and BAX polymerization by MCL-1 is prevented either by MCL-1 degradation or by saturating and inhibiting the MCL-1 binding sites on BAK/BAX with BH3 proteins or mimetics.Under normal growth conditions, MCL-1 is important for mouse embryonic survival (4) and critical for the survival of neutrophils, lymphocytes, hematopoietic stem cells, and neurons (5). MCL-1 overexpression is the hallmark of several cancers, including hematological malignancies as well as solid tumors. Elevated cellular MCL-1 expression correlates with resistance to drug toxicity and ionizing radiation (IR), whereas its inhibition sensitizes cells to both. The BCL-2 family of proteins is characterized by the presence of BCL-2 homology (BH) domains (1, 2). The MCL-1 protein itself is unique among BCL-2 members in also containing multiple N-terminal PEST motifs in addition to BH1, BH2, BH3, and C-terminal transmembrane (TM) domains. PEST is ...

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Section: Mcl-1 In Dna Damage Repair Molecular and Cellular Biologymentioning

confidence: 99%

MCL-1 Depletion Impairs DNA Double-Strand Break Repair and Reinitiation of Stalled DNA Replication Forks

Mattoo

Pandita

Chakraborty

et al. 2017

Molecular and Cellular Biology

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“…αII Sp functions together with the DNA ICL repair proteins XPF and FANCA to localize at nuclear foci after DNA ICL damage [ 16 , 18 , 20 ]. A recent report has shown β2 spectrin deficiency disturbs chromosome stability [ 21 ], and a number of studies have shown that spectrin is an essential regulator in a variety of cancers [ 22 – 26 ].…”

Section: Introductionmentioning

confidence: 99%

MicroRNA-128-3p regulates mitomycin C-induced DNA damage response in lung cancer cells through repressingSPTAN1

et al. 2016

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The DNA damage response is critical for maintaining genome integrity and preventing damage to DNA due to endogenous and exogenous insults. Mitomycin C (MMC), a potent DNA cross-linker, is used as a chemotherapeutic agent because it causes DNA inter-strand cross-links (DNA ICLs) in cancer cells. While many microRNAs, which may serve as oncogenes or tumor suppressors, are grossly dysregulated in human cancers, little is known about their roles in MMC-treated lung cancer. Here, we report that miR-128-3p can attenuate repair of DNA ICLs by targeting SPTAN1 (αII Sp), resulting in cell cycle arrest and promoting chromosomal aberrations in lung cancer cells treated with MMC. Using computational prediction and experimental validation, SPTAN1 was found to be a conserved target of miR-128-3p. We then found that miR-128-3p caused translational inhibition of SPTAN1, reducing its protein level. SPTAN1 repression via miR-128-3p also induced cell cycle arrest and chromosomal instability. Additionally, miR-128-3p significantly influenced interaction of the αII Sp/FANCA/XPF complex, thus limiting DNA repair. In summary, the results demonstrate that miR-128-3p accelerates cell cycle arrest and chromosomal instability in MMC-treated lung cancer cells by suppressing SPTAN1, and these findings could be applied for adjuvant chemotherapy of lung cancer.

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“…for 30 mins before stalling with 2mM Hydroxy Urea (HU) for 2h,then washed in PBS for 2 times, incubated in medium containing with 450 mM Iodoxy Uridine (IdU) for 1hr Horikoshi et al, 2016).…”

Section: Dna Replication Restart Assay and Fork Protection Assaymentioning

confidence: 99%

Role of HP1β during spermatogenesis and DNA replication

Charakha¹,

Tiwari

Pandita

et al. 2019

Preprint

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Maintaining genomic stability in a continually dividing cell population requires accurate DNA repair, especially in male germ cells. Repair and replication protein access to DNA, however, is complicated by chromatin compaction. The HP1β chromatin protein, encoded by Cbx1, is associated with chromatin condensation but its role in meiosis is not clear. To investigate the role of Cbx1 in male germ cells, we generated testis specific Cbx1 deficient transgenic mice by crossing Cbx1 flox/flox (Cbx1 f/f ) mice with Stra8 Cre +/mice. Loss of Cbx1 in testes adversely affected sperm maturation and Cbx1 deletion increased seminiferous tubule degeneration and basal level DNA damage., We observed that Cbx1 -/-MEF cells displayed reduced resolution of stalled DNA replication forks as well as decreased fork restart, indicating defective DNA synthesis. Taken together, these results suggest that loss of Cbx1 in growing cells leads to DNA replication defects and associated DNA damage that impact cell survival.

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β2-spectrin depletion impairs DNA damage repair

Cited by 19 publications

References 64 publications

MCL-1 Depletion Impairs DNA Double-Strand Break Repair and Reinitiation of Stalled DNA Replication Forks

MCL-1 Depletion Impairs DNA Double-Strand Break Repair and Reinitiation of Stalled DNA Replication Forks

MicroRNA-128-3p regulates mitomycin C-induced DNA damage response in lung cancer cells through repressingSPTAN1

Role of HP1β during spermatogenesis and DNA replication

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