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In late mitosis and early G1, replication origins are licensed for subsequent use by loading complexes of the minichromosome maintenance proteins 2-7 (Mcm2-7). The number of Mcm2-7 complexes loaded onto DNA greatly exceeds the number of replication origins used during S phase, but the function of the excess Mcm2-7 is unknown. Using Xenopus laevis egg(More)
Accurate DNA replication requires proper regulation of replication licensing, which entails loading MCM-2-7 onto replication origins. In this paper, we provide the first comprehensive view of replication licensing in vivo, using video microscopy of Caenorhabditis elegans embryos. As expected, MCM-2-7 loading in late M phase depended on the prereplicative(More)
Before S phase, cells license replication origins for initiation by loading them with Mcm2-7 heterohexamers. This process is dependent on Cdc6, which is recruited to unlicensed origins. Using Xenopus egg extracts we show that although each origin can load many Mcm2-7 hexamers, the affinity of Cdc6 for each origins drops once it has been licensed by loading(More)
Holliday junctions (HJs) are cruciform DNA structures that are created during recombination events. It is a matter of considerable importance to determine the resolvase(s) that promote resolution of these structures. We previously reported that C. elegans GEN-1 is a symmetrically cleaving HJ resolving enzyme required for recombinational repair, but we could(More)
Meiotic recombination is essential for the repair of programmed double strand breaks (DSBs) to generate crossovers (COs) during meiosis. The efficient processing of meiotic recombination intermediates not only needs various resolvases but also requires proper meiotic chromosome structure. The Smc5/6 complex belongs to the structural maintenance of(More)
The small ubiquitin-like modifier (SUMO), initially characterized as a suppressor of a mutation in the gene encoding the centromeric protein MIF2, is involved in many aspects of cell cycle regulation. The dynamics of conjugation and deconjugation and the role of SUMO during the cell cycle remain unexplored. Here we used Caenorhabditis elegans to establish(More)
This Article contains errors in the legends to Supplementary Movies 9–11, which attribute fluorescence to cells expressing mCherry-H2B/GFP-AIR-2. These cells express mCherry-SMO-1(GG)/GFP-H2B. There is also an error in Fig. 1e that was introduced during the production process and resulted in the error bars for the 100 s time point being displaced to an(More)
Expression and purification of non-degradable geminin DEL were described in [S1]. Mcm2, Mcm3, Smc2 ,and Nup153 antibodies were as described previously [S2]. Rabbit polyclonal antiserum was raised against a synthetic peptide corresponding to the alpha-peptide of Xenopus pol d (GTKQA SIMGFFQKK) as described [S3]. Xenopus Chromatin Manipulations Mitotically(More)
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