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Dual DNA replication modes: varying fork speeds and initiation rates within the spatial replication program in Xenopus.
Nucleic Acids Res
January 2025
Université Paris Cité, CNRS, Institut Jacques Monod, F-75013 Paris, France.
Large vertebrate genomes duplicate by activating tens of thousands of DNA replication origins, irregularly spaced along the genome. The spatial and temporal regulation of the replication process is not yet fully understood. To investigate the DNA replication dynamics, we developed a methodology called RepliCorr, which uses the spatial correlation between replication patterns observed on stretched single-molecule DNA obtained by either DNA combing or high-throughput optical mapping.
View Article and Find Full Text PDFCryo-EM structure of AAV2 Rep68 bound to integration site AAVS1: insights into the mechanism of DNA melting.
Nucleic Acids Res
January 2025
Department of Physiology and Biophysics, Virginia Commonwealth University, School of Medicine, Richmond, VA 23298, United States.
The Rep68 protein from Adeno-Associated Virus (AAV) is a multifunctional SF3 helicase that performs most of the DNA transactions necessary for the viral life cycle. During AAV DNA replication, Rep68 assembles at the origin of replication, catalyzing the DNA melting and nicking reactions during the hairpin rolling replication process to complete the second-strand synthesis of the AAV genome. We report the cryo-electron microscopy structures of Rep68 bound to the adeno-associated virus integration site 1 in different nucleotide-bound states.
View Article and Find Full Text PDFReplication across O6-methylguanine activates futile cycling of DNA mismatch repair attempts assisted by the chromatin remodeling enzyme Smarcad1.
J Biochem
January 2025
Faculty of Science, Kyushu University, 744 Motooka, Nishi-ku, Fukuoka 819-0395, Japan.
SN1-type alkylating reagents generate O6-methylguanine (meG) lesions that activate the mismatch repair (MMR) response. Since post-replicative MMR specifically targets the nascent strand, meG on the template strand is refractory to rectification by MMR and, therefore, can induce non-productive MMR reactions. The cycling of futile MMR attempts is proposed to cause DNA double-strand breaks in the subsequent S phase, leading to ATR-checkpoint-mediated G2 arrest and apoptosis.
View Article and Find Full Text PDFA yeast-based reverse genetics system to generate HCoV-OC43 reporter viruses encoding an eighth subgenomic RNA.
J Virol
January 2025
Department of Microbiology and Immunology, Dalhousie University, Halifax, Nova Scotia, Canada.
Unlabelled: Coronaviruses have large, positive-sense single-stranded RNA genomes that challenge conventional strategies for mutagenesis. Yeast genetics has been used to manipulate large viral genomes, including those of herpesviruses and coronaviruses. This method, known as transformation-associated recombination (TAR), involves assembling complete viral genomes from dsDNA copies of viral genome fragments via homologous recombination in .
View Article and Find Full Text PDFChloroplast State Transitions Modulate Nuclear Genome Stability via Cytokinin Signaling in Arabidopsis.
Mol Plant
January 2025
Institute of Medical Plant Physiology and Ecology, School of Pharmaceutical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, 510006 People's Republic of China; State Key Laboratory of Traditional Chinese Medicine Syndrome, Guangzhou University of Chinese Medicine, Guangzhou, 510006, People's Republic of China; Chinese Medicine Guangdong Laboratory, Guangdong Hengqin, People's Republic of China. Electronic address:
Activities of the chloroplasts and nucleus are coordinated by retrograde signaling, which has crucial roles in plant development and environmental adaptation. However, the connection between chloroplast status and nuclear genome stability is not well understood. Chloroplast state transitions allow the plant to balance the absorption capacity of the photosystems in an environment in which the light quality was changing.
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