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doi: 10.4161/cc.19709
pmid: 22421151
Several replication-initiation proteins are assembled stepwise onto replicators to form pre-replicative complexes (pre-RCs) to license eukaryotic DNA replication. We performed a yeast functional proteomic screen and identified the Rix1 complex members (Ipi1p-Ipi2p/Rix1-Ipi3p) as pre-RC components and critical determinants of replication licensing and replication-initiation frequency. Ipi3p interacts with pre-RC proteins, binds chromatin predominantly at ARS sequences in a cell cycle-regulated and ORC- and Noc3p-dependent manner and is required for loading Cdc6p, Cdt1p and MCM onto chromatin to form pre-RC during the M-to-G₁ transition and for pre-RC maintenance in G₁ phase-independent of its role in ribosome biogenesis. Moreover, Ipi1p and Ipi2p, but not other ribosome biogenesis proteins Rea1p and Utp1p, are also required for pre-RC formation and maintenance, and Ipi1p, -2p and -3p are interdependent for their chromatin association and function in pre-RC formation. These results establish a new framework for the hierarchy of pre-RC proteins, where the Ipi1p-2p-3p complex provides a critical link between ORC-Noc3p and Cdc6p-Cdt1p-MCM in replication licensing.
DNA Replication, 570, Nucleocytoplasmic Transport Proteins, Replication licensing, Saccharomyces cerevisiae Proteins, Proteome, Ipi2p and Ipi3p) complex, Origin Recognition Complex, Cell Cycle Proteins, Replication Origin, Saccharomyces cerevisiae, DNA replication, Pre-replicative complex, Cell Cycle, DNA Helicases, G1 Phase, Nuclear Proteins, Minichromosome Maintenance 1 Protein, 540, Chromatin, DNA-Binding Proteins, Trans-Activators, The Rix 1 (Ipi1p, Ribosomes
DNA Replication, 570, Nucleocytoplasmic Transport Proteins, Replication licensing, Saccharomyces cerevisiae Proteins, Proteome, Ipi2p and Ipi3p) complex, Origin Recognition Complex, Cell Cycle Proteins, Replication Origin, Saccharomyces cerevisiae, DNA replication, Pre-replicative complex, Cell Cycle, DNA Helicases, G1 Phase, Nuclear Proteins, Minichromosome Maintenance 1 Protein, 540, Chromatin, DNA-Binding Proteins, Trans-Activators, The Rix 1 (Ipi1p, Ribosomes
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