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Key Points Activation of initiation of DNA replication occurs at only a subset of replication origins that were previously assembled in the G1 phase of the cell cycle. This is achieved through a highly regulated sequential two-step process: origin licensing in the G1 phase and origin activation during the S phase. DNA replication origins from Saccharomyces cerevisiae have a sequence consensus, whereas metazoan origins are more plastic and are determined by both sequence preferences, such as G-rich elements, and epigenetic features. Chromosomal environment and transcriptional status also influence the activation of origins. Origins selected in adjacent replication units are synchronously activated and form replication domains, which are activated at specific times during the S phase. Replication timing domains correlate with topologically associated domains (TADs). Replication timing is regulated by specific proteins and chromatin marks. The activation of DNA replication origins is developmentally controlled. Specific checkpoints regulate the initiation of DNA replication in response to replication stress. During the G1–S phase transition of the cell cycle, a variable subset of previously 'licensed' origins of replication is activated to initiate DNA synthesis. Insight is being gained into the mechanisms underlying which origins are activated and when; these mechanisms are associated with nuclear organization, cell differentiation and replication stress. DNA replication begins with the assembly of pre-replication complexes (pre-RCs) at thousands of DNA replication origins during the G1 phase of the cell cycle. At the G1–S-phase transition, pre-RCs are converted into pre-initiation complexes, in which the replicative helicase is activated, leading to DNA unwinding and initiation of DNA synthesis. However, only a subset of origins are activated during any S phase. Recent insights into the mechanisms underlying this choice reveal how flexibility in origin usage and temporal activation are linked to chromosome structure and organization, cell growth and differentiation, and replication stress.

Cell death occurring during mitosis, or mitotic catastrophe, often takes place in conjunction with apoptosis, but the conditions in which mitotic catastrophe may exhibit features of programmed cell death are still unclear. In the work presented here, we studied mitotic cell death by making use of a UV-inactivated parvovirus (adeno-associated virus; AAV) that has been shown to induce a DNA damage response and subsequent death of p53-defective cells in mitosis, without affecting the integrity of the host genome. Osteosarcoma cells (U2OSp53DD) that are deficient in p53 and lack the G1 cell cycle checkpoint respond to AAV infection through a transient G2 arrest. We found that the infected U2OSp53DD cells died through mitotic catastrophe with no signs of chromosome condensation or DNA fragmentation. Moreover, cell death was independent of caspases, apoptosis-inducing factor (AIF), autophagy and necroptosis. These findings were confirmed by time-lapse microscopy of cellular morphology following AAV infection. The assays used readily revealed apoptosis in other cell types when it was indeed occurring. Taken together the results indicate that in the absence of the G1 checkpoint, mitotic catastrophe occurs in these p53-null cells predominantly as a result of mechanical disruption induced by centrosome overduplication, and not as a consequence of a suicide signal.

Upon fertilisation, the sperm pronucleus acquires the competence to replicate the genome through a cascade of events that link chromatin remodelling to nuclear envelope formation. The factors involved have been partially identified and are poorly characterised. Here, using Xenopus laevis egg extracts we show that RNAs are required for proper nuclear envelope assembly following sperm DNA decondensation. Although chromatin remodelling and pre-replication complex formation occur normally, RNA-depleted extracts show a defect in pre-RC activation. The nuclear processes affected by RNA-depletion included ELYS recruitment, which accounts for the deficiency in nuclear pore complex assembly. This results in failure in chromatin relaxation as well as in the import and proper nuclear concentration of the S-phase kinases necessary for DNA replication activation. Our results highlight a translation-independent RNA function necessary for the parental genome progression towards the early embryonic cell cycle programme. The factors that link chromatin remodelling to nuclear envelope formation in the sperm pronucleus are not fully characterised. Here, the authors show that in RNA-depleted Xenopus laevis egg extracts, ELYS recruitment and nuclear pore complex formation are impaired, resulting in defective nuclear processes.

In the original version of this Article, the affiliation details for Antoine Aze, Michalis Fragkos, Stéphane Bocquet, Julien Cau and Marcel Méchali incorrectly omitted ‘CNRS and the University of Montpellier’. This has now been corrected in both the PDF and HTML versions of the Article.

Gene therapy applications that target hematopoietic stem cells (HSCs) offer great potential for the treatment of hematologic disease. Despite this promise, clinical success has been limited by poor rates of gene transfer, poor engraftment of modified cells, and poor levels of gene expression. We describe here the basic approach used for HSC gene therapy, briefly review some of the seminal clinical trials in the field, and describe several recent advances directed toward overcoming these limitations.

Cancer is a condition characterized by genomic instability and gross chromosomal aberrations. The inability of the cell to timely and efficiently complete its replication cycle before entering mitosis is one of the most common causes of DNA damage and carcinogenesis. Phosphorylation of histone 2AX (H2AX) on S139 (γH2AX) is an indispensable step in the response to DNA damage, as it is required for the assembly of repair factors at the sites of damage. γH2AX is also a marker of DNA replication stress, mainly due to fork collapse that often follows prolonged replication stalling or repair of arrested forks, which involves the generation of DNA breaks. Although the role of γH2AX in the repair of DNA breaks has been well defined, the function of γH2AX in replicative stress remains unclear. In this review, we present the recent advances in the field of replication stress, and highlight a novel function for γH2AX that is independent of its role in the response to DNA damage. We discuss studies that support a role for γΗ2ΑΧ early in the response to replicative stress, which does not involve the repair of DNA breaks. We also highlight recent data proposing that γH2AX acts as a chromatin remodeling component, implicated in the efficient resolution of stalled replication forks. Understanding the mechanism by which γH2AX enables cellular recovery after replication stress will allow identification of novel cancer biomarkers, as well as new targets for cancer therapies.

Earth observation (EO) techniques have significantly evolved over time, covering a wide range of applications in different domains. The scope of this study is to review the research conducted on EO in the Eastern Mediterranean, Middle East, and North Africa (EMMENA) region and to identify the main knowledge gaps. We searched through the Web of Science database for papers published between 2018 and 2022 for EO studies in the EMMENA. We categorized the papers in the following thematic areas: atmosphere, water, agriculture, land, disaster risk reduction (DRR), cultural heritage, energy, marine safety and security (MSS), and big Earth data (BED); 6647 papers were found with the highest number of publications in the thematic areas of BED (27%) and land (22%). Most of the EMMENA countries are surrounded by sea, yet there was a very small number of studies on MSS (0.9% of total number of papers). This study detected a gap in fundamental research in the BED thematic area. Other future needs identified by this study are the limited availability of very high-resolution and near-real-time remote sensing data, the lack of harmonized methodologies and the need for further development of models, algorithms, early warning systems, and services.