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Principal Investigator: Franziska Bleichert
Organization: YALE UNIVERSITY
Fiscal Year: 2024
Award: $321,155
Funding agency: National Institute of General Medical Sciences
Project Summary
The survival of living organisms depends on the timely and accurate duplication of chromosomal DNA. In all
domains of life, the onset of DNA replication (or initiation) relies on dedicated initiator proteins that bind
genomic sites, termed replication origins, and help load replicative helicases onto DNA. In eukaryotes, the
initiator is the origin recognition complex (ORC), which recruits and deposits the Mcm2-7 helicase motor
module onto DNA to ‘license’ origins. Although core replication initiation factors are conserved across most
eukaryotes and are well-studied in budding yeast, important differences exist with respect to origin recognition
and the regulation of origin licensing between S. cerevisiae and higher eukaryotes. Numerous outstanding
questions therefore remain in the metazoan system regarding origin recruitment and function of core and
accessory DNA replication initiation factors, and the specific contributions of chromatin-associated proteins to
these events. In this proposal, we aim to define at a mechanistic level in metazoan systems how ORC-
dependent DNA remodeling contributes to Mcm2-7 loading, how disease-linked mutations alter initiator
activities, and how ORC-partner proteins promote chromosomal recruitment and function of the initiator, by
integrating biochemical, structural, and cell-based approaches. The findings from this work will have broad
implications for multiple scientific fields, as they will not only help generate models for origin specification,
origin processing, and origin licensing in metazoans, but also contribute to our understanding of DNA- and
chromatin-dependent macromolecular machines. Our studies are also of significant biomedical relevance since
the failure to precisely replicate chromosomal DNA causes genome instability, which in turn underpins many
human diseases, including cancer. In the long-term, the outcomes of our efforts thus have the potential to
provide important starting points for the development of novel therapeutic strategies.
Terms: <Address><Assay><Baker's Yeast><Basal Cell Nevus Syndrome><Binding><Binding Proteins><Bioassay><Biochemical><Biological Assay><Brewer's Yeast><Budding Yeast><Cancers><Cell Body><Cell Growth in Number><Cell Multiplication><Cell Proliferation><Cell Survival><Cell Viability><Cells><Cellular Expansion><Cellular Growth><Cellular Proliferation><Chromatin><Chromosomal Amplification><Chromosomal Duplication><Complex><Cryo-electron Microscopy><Cryoelectron Microscopy><DNA><DNA Binding><DNA Binding Interaction><DNA Helicases><DNA Replication><DNA Replication Initiation><DNA Sequence><DNA Synthesis><DNA Unwinding Proteins><DNA biosynthesis><DNA bound><DNA unwinding enzyme><Data><Dedications><Deoxyribonucleic Acid><Deposit><Deposition><Development><Disease><Disorder><Dwarfism><Electron Cryomicroscopy><Endomycetales><Environment><Eukaryota><Eukaryote><Event><Failure><Fifth Phacomatosis><Generalized Growth><Genetic Alteration><Genetic Change><Genetic defect><Genome Instability><Genomic Instability><Genomics><Goals><Gorlin Syndrome><Gorlin syndrome 2><Gorlin-Goltz Syndrome><Growth><In Vitro><Initiation Factors><Knowledge><Label><Laboratories><Learning><Licensing><Life><Ligand Binding Protein><Ligand Binding Protein Gene><Link><Maintenance><Malignant Neoplasms><Malignant Tumor><Mediating><Modeling><Molecular><Molecular Interaction><Motor><Mutation><Nanism><Nevoid Basal Cell Carcinoma Syndrome><Nucleosomes><ORC protein><Organism><Outcome><Peptide Initiation Factors><Position><Positioning Attribute><Process><Protein Binding><Proteins><Regulation><Replication Initiation><Replication Origin><Research><Research Proposals><S cerevisiae><S. cerevisiae><Saccharomyces cerevisiae><Saccharomycetales><Site><Specific qualifier value><Specified><Structure><System><Techniques><Testing><Tissue Growth><Translation Initiation Factor><Translational Initiation Factor><Work><Yeast Model System><biochemical model><bound protein><cell growth><cofactor><cryo-EM><cryoEM><cryogenic electron microscopy><developmental><dwarf><genetic information><genome mutation><helicase><hereditary cutaneomandibular polyoncosis><human disease><in vivo><jaw cysts-basal cell tumors-skeletal anomalies syndrome><living system><malignancy><multiple basal-cell carcinoma syndrome multiple basal-cell nevus syndrome><multiple hereditary cutaneomandibular polyoncosis><multiple nevoid basal-cell carcinoma syndrome><multiple nevoid basal-cell epithelioma-jaw cysts-bifid rib syndrome><neoplasm/cancer><nevoid basal-cell epithelioma-jaw cysts-bifid rib syndrome><new therapeutic approach><new therapeutic intervention><new therapeutic strategies><new therapy approaches><new treatment approach><new treatment strategy><novel><novel therapeutic approach><novel therapeutic intervention><novel therapeutic strategies><novel therapy approach><odontogenic keratocytosis-skeletal anomalies syndrome><ontogeny><ori Region><origin recognition complex><recruit><yeast model>