Chromatin bridges: stochastic breakage or regulated resolution?

Genetic material is organized in the form of chromosomes, which need to be segregated accurately into two daughter cells in each cell cycle. However, chromosome fusion or the presence of unresolved interchromosomal linkages lead to the formation of chromatin bridges, which can induce DNA lesions and genome instability. Persistent chromatin bridges are trapped in the cleavage furrow and are broken at or after abscission, the final step of cytokinesis. In this review, we focus on recent progress in understanding the mechanism of bridge breakage and resolution. We discuss the molecular machinery and enzymes that have been implicated in the breakage and processing of bridge DNA. In addition, we outline both the immediate outcomes and genomic consequences induced by bridge breakage.

Human Endonuclease ANKLE1 Localizes at the Midbody and Processes Chromatin Bridges to Prevent DNA Damage and cGAS-STING Activation.

Chromatin bridges connecting the two segregating daughter nuclei arise from chromosome fusion or unresolved interchromosomal linkage. Persistent chromatin bridges are trapped in the cleavage plane, triggering cytokinesis delay. The trapped bridges occasionally break during cytokinesis, inducing DNA damage and chromosomal rearrangements. Recently, Caenorhabditis elegans LEM-3 and human TREX1 nucleases have been shown to process chromatin bridges. Here, it is shown that ANKLE1 endonuclease, the human ortholog of LEM-3, accumulates at the bulge-like structure of the midbody via its N-terminal ankyrin repeats. Importantly, ANKLE1-/- knockout cells display an elevated level of G1-specific 53BP1 nuclear bodies, prolonged activation of the DNA damage response, and replication stress. Increased DNA damage observed in ANKLE1-/- cells is rescued by inhibiting actin polymerization or reducing actomyosin contractility. ANKLE1 does not act in conjunction with structure-selective endonucleases, GEN1 and MUS81 in resolving recombination intermediates. Instead, ANKLE1 acts on chromatin bridges by priming TREX1 nucleolytic activity and cleaving bridge DNA to prevent the formation of micronuclei and cytosolic dsDNA that activate the cGAS-STING pathway. It is therefore proposed that ANKLE1 prevents DNA damage and autoimmunity by cleaving chromatin bridges to avoid catastrophic breakage mediated by actomyosin contractile forces.

Identification and characterization of a novel gene, c1orf109, encoding a CK2 substrate that is involved in cancer cell proliferation.

BACKGROUND: In the present study we identified a novel gene, Homo Sapiens Chromosome 1 ORF109 (c1orf109, GenBank ID: NM_017850.1), which encodes a substrate of CK2. We analyzed the regulation mode of the gene, the expression pattern and subcellular localization of the predicted protein in the cell, and its role involving in cell proliferation and cell cycle control. METHODS: Dual-luciferase reporter assay, chromatin immunoprecipitation and EMSA were used to analysis the basal transcriptional requirements of the predicted promoter regions. C1ORF109 expression was assessed by western blot analysis. The subcellular localization of C1ORF109 was detected by immunofluorescence and immune colloidal gold technique. Cell proliferation was evaluated using MTT assay and colony-forming assay. RESULTS: We found that two cis-acting elements within the crucial region of the c1orf109 promoter, one TATA box and one CAAT box, are required for maximal transcription of the c1orf109 gene. The 5' flanking region of the c1orf109 gene could bind specific transcription factors and Sp1 may be one of them. Employing western blot analysis, we detected upregulated expression of c1orf109 in multiple cancer cell lines. The protein C1ORF109 was mainly located in the nucleus and cytoplasm. Moreover, we also found that C1ORF109 was a phosphoprotein in vivo and could be phosphorylated by the protein kinase CK2 in vitro. Exogenous expression of C1ORF109 in breast cancer Hs578T cells induced an increase in colony number and cell proliferation. A concomitant rise in levels of PCNA (proliferating cell nuclear antigen) and cyclinD1 expression was observed. Meanwhile, knockdown of c1orf109 by siRNA in breast cancer MDA-MB-231 cells confirmed the role of c1orf109 in proliferation. CONCLUSIONS: Taken together, our findings suggest that C1ORF109 may be the downstream target of protein kinase CK2 and involved in the regulation of cancer cell proliferation.

Puerarin attenuates preeclampsia-induced trophoblast mobility loss and inflammation by modulating miR-181b-5p/RBAK axis.

PROBLEM: Preeclampsia (PE) is a serious pregnancy complication characterized by inflammation and impaired trophoblast motility. Puerarin (Pue) is a functional compound with anti-PE potential. The current study aimed to explore the therapeutic effects of Pue on PE as well as the associated mechanism by focusing on the interaction between Pue and microRNAs (miRs). METHODS OF STUDY: Human villous trophoblast HTR-8/SVneo cells were treated with TNF-α and Pue, and a change in miR expression profile was determined. The anti-PE effects of Pue were validated in rat models by measuring blood pressure, 24-h proteinuria, and cytokine levels. The mechanism was explored by focusing on miR-181b-5p/RBAK axis. RESULTS: The induction of PE increased blood pressure and 24-h proteinuria, and induced TNF-α, IL-1ß, and IL-6 levels, which was reversed by Pue. In in vitro assays, TNF-α suppressed viability, induced apoptosis and inflammatory response, and inhibited migration in trophoblasts, which was attenuated by Pue. At molecular level, the expression level of miR-181b-5p was both induced in vivo and in vitro with the development of PE symptoms, contributing to the inhibited expression of RBAK. The induced expression of miR-181b-5p under Pue treatment showed that the reinduction of miR-181b-5p counteracted the effects of Pue, indicating the key role of the miR in the protective effects of Pue against PE. CONCLUSIONS: The current study verified the anti-PE function of Pue: the compound suppressed inflammatory response associated with PE, and improved trophoblast motility. The effects depended on the inhibition of miR-181b-5p that inhibited the expression of RBAK.

Polyvinylidene fluoride multi-scale nanofibrous membrane modified using N-halamine with high filtration efficiency and durable antibacterial properties for air filtration.

HYPOTHESIS: Particulate matter (PM) pollution and the coronavirus (COVID-19) pandemic have increased demand for protective masks. However, typical protective masks only intercept particles and produce peculiar odors if worn for extended periods owing to bacterial growth. Therefore, new protective materials with good filtration and antibacterial capabilities are required. EXPERIMENTS: In this study, we prepared multi-scale polyvinylidene fluoride (PVDF) nanofibrous membranes for efficient filtration and durable antibacterial properties via N-halamine modification. FINDINGS: The N-halamine-modified nanofibrous membrane (PVDF-PAA-TMP-Cl) had sufficient active chlorine content (800 ppm), and the tensile stress and strain were improved compared with the original membrane, from 6.282 to 9.435 MPa and from 51.3 % to 56.4 %, respectively. To further improve the interception efficiency, ultrafine nanofibers (20-35 nm) were spun on PVDF-PAA-TMP-Cl nanofibrous membranes, and multi-scale PVDF-PAA-TMP-Cl nanofibrous membranes were prepared. These membranes exhibited good PM0.3 interception (99.93 %), low air resistance (79 Pa), promising long-term PM2.5 purification ability, and high bactericidal efficiency (>98 %). After ten chlorination cycles, the antibacterial efficiency against Escherichia coli and Staphylococcus aureus exceeded 90 %; hence, the material demonstrated highly efficient filtration and repeatable antibacterial properties. The results of this study have implications for the development of air and water filtration systems and multi-functional protective materials.

[Reduction of selenite to elemental red selenium under aerobic condition by Pseudomonas alcaliphila MBR].

OBJECTIVE: We studied the reduction of soluble and highly toxic selenite to elemental red selenium, under aerobic condition by Pseudomonas alcaliphila MBR, with organic carbon as electron donor. RESULTS: The strain could grow under pH 6-11 and resist to high concentration of selenite with the minimal inhibitory concentration of 50 mmol/L. After 5 days, the strain used sodium citrate as electron donor, and reduced 2.0 mmol/L selenite to elemental red selenium from the culture fluid, the elemental red selenium was stored outside the cells. The glutathione and nitrate could increase the number of reduction rate. CONCLUSION: This study implies the application of Pseudomonas alcaliphila MBR to convert selenite to elemental red selenium.

FOP Negatively Regulates Ciliogenesis and Promotes Cell Cycle Re-entry by Facilitating Primary Cilia Disassembly.

Primary cilia are microtubule-based, antenna-like organelles, which are formed in G0 phase and resorbed as cells re-enter the cell cycle. It has been reported that primary cilia can influence the timing of cell cycle progression. However, the molecular links between ciliogenesis and cell cycle progression are not well understood. The Fibroblast Growth Factor Receptor 1 Oncogene Partner (FOP) has been implicated in ciliogenesis, but its function in ciliogenesis is not clear. Here, we show that FOP plays a negative role in ciliogenesis. Knockdown of FOP promotes cilia elongation and suppresses cilia disassembly. In contrast, ectopic expression of FOP induces defects in primary cilia formation, which can be rescued by either pharmacological or genetic inhibition of Aurora kinase A which promotes cilia disassembly. Moreover, knockdown of FOP delays cell cycle re-entry of quiescent cells following serum re-stimulation, and this can be reversed by silencing Intraflagellar Transport 20 (IFT20), an intraflagellar transport member essential for ciliogenesis. Collectively, these results suggest that FOP negatively regulates ciliogenesis and can promote cell cycle re-entry by facilitating cilia disassembly.

A Role of hIPI3 in DNA Replication Licensing in Human Cells.

The yeast Ipi3p is required for DNA replication and cell viability in Sacharomyces cerevisiae. It is an essential component of the Rix1 complex (Rix1p/Ipi2p-Ipi1p-Ipi3p) that is required for the processing of 35S pre-rRNA in pre-60S ribosomal particles and for the initiation of DNA replication. The human IPI3 homolog is WDR18 (WD repeat domain 18), which shares significant homology with yIpi3p. Here we report that knockdown of hIPI3 resulted in substantial defects in the chromatin association of the MCM complex, DNA replication, cell cycle progression and cell proliferation. Importantly, hIPI3 silencing did not result in a reduction of the protein level of hCDC6, hMCM7, or the ectopically expressed GFP protein, indicating that protein synthesis was not defective in the same time frame of the DNA replication and cell cycle defects. Furthermore, the mRNA and protein levels of hIPI3 fluctuate in the cell cycle, with the highest levels from M phase to early G1 phase, similar to other pre-replicative (pre-RC) proteins. Moreover, hIPI3 interacts with other replication-initiation proteins, co-localizes with hMCM7 in the nucleus, and is important for the nuclear localization of hMCM7. We also found that hIPI3 preferentially binds to the origins of DNA replication including those at the c-Myc, Lamin-B2 and ß-Globin loci. These results indicate that hIPI3 is involved in human DNA replication licensing independent of its role in ribosome biogenesis.

[Observation of the metatarsal nutrient artery]

OBJECTIVE: To observe the origin, course and diameter of the metatarsal nutrient artery. METHODS: The metatarsaL nutrient in 90 feet, ranging in age from newborn to 87 years, were studied by perfusion method. The origin, course and diameter of these arteries were observed and measured. RESULTS: The diameter of the metatarsal nutrient artery was 0.24 approximate, equals 0.30 mm. The nutrient arteries of the first metatarsal bone originated from the deep plantar branch and the first metatarsal plantar artery in 59.6% of specimens, while the nutrient arteries of the other metatarsal bones mainly originate from the plantar metatarsal arteries, the plantar arch and its perfora-ting branches. The diaphysial nutrient foramina were situated in the middle third of the shaft over 90% of specimens. CONCLUSION: The metatarsal nutrient artery showes practical significance in clinic.

AIF downregulation and its interaction with STK3 in renal cell carcinoma.

Apoptosis-inducing factor (AIF) plays a crucial role in caspase-independent programmed cell death by triggering chromatin condensation and DNA fragmentation. Therefore, it might be involved in cell homeostasis and tumor development. In this study, we report significant AIF downregulation in the majority of renal cell carcinomas (RCC). In a group of RCC specimens, 84% (43 out of 51) had AIF downregulation by immunohistochemistry stain. Additional 10 kidney tumors, including an oxyphilic adenoma, also had significant AIF downregulation by Northern blot analysis. The mechanisms of the AIF downregulation included both AIF deletion and its promoter methylation. Forced expression of AIF in RCC cell lines induced massive apoptosis. Further analysis revealed that AIF interacted with STK3, a known regulator of apoptosis, and enhanced its phosphorylation at Thr180. These results suggest that AIF downregulation is a common event in kidney tumor development. AIF loss may lead to decreased STK3 activity, defective apoptosis and malignant transformation.