Replication foci dynamics: replication patterns are modulated by S-phase checkpoint kinases in fission yeast

Meister, Peter; Taddei, Angela; Ponti, Aaron; Baldacci, Giuseppe; Gasser, Susan M.

doi:10.1038/sj.emboj.7601538

Cited by 78 publications

(92 citation statements)

References 47 publications

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“…The pattern of these foci changes with S-phase progression. While a greater number of foci are apparent in early and mid S phase, fewer foci are seen in late S phase (Meister et al, 2007). These changes are believed to mirror the pattern of DNA synthesis in S phase, with transcriptionally active euchromatin replicated early in S phase, and heterochromatic DNA replicated later on.…”

Section: Pcna Is Nuclear Throughout the Cell Cycle But Is Mainly Expmentioning

confidence: 97%

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The distribution pattern of proliferating cell nuclear antigen in the nuclei of Leishmania donovani

et al. 2009

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DNA replication in eukaryotes is a highly conserved process marked by the licensing of multiple origins, with pre-replication complex assembly in G1 phase, followed by the onset of replication at these origins in S phase. The two strands replicate by different mechanisms, and DNA synthesis is brought about by the activity of the replicative DNA polymerases Pol d and Pol e. Proliferating cell nuclear antigen (PCNA) augments the processivity of these polymerases by serving as a DNA sliding clamp protein. This study reports the cloning of PCNA from the protozoan Leishmania donovani, which is the causative agent of the systemic disease visceral leishmaniasis. PCNA was demonstrated to be robustly expressed in actively proliferating L. donovani promastigotes. We found that the protein was present primarily in the nucleus throughout the cell cycle, and it was found in both proliferating procyclic and metacyclic promastigotes. However, levels of expression of PCNA varied through cell cycle progression, with maximum expression evident in G1 and S phases. The subnuclear pattern of expression of PCNA differed in different stages of the cell cycle; it formed distinct subnuclear foci in S phase, while it was distributed in a more diffuse pattern in G2/M phase and post-mitotic phase cells. These subnuclear foci are the sites of active DNA replication, suggesting that replication factories exist in Leishmania, as they do in higher eukaryotes, thus opening avenues for investigating other Leishmania proteins that are involved in DNA replication as part of these replication factories.

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Section: Pcna Is Nuclear Throughout the Cell Cycle But Is Mainly Expmentioning

confidence: 97%

“…The dynamics of these foci have been studied in Schizosaccharomyces pombe and mammalian cells (Leonhardt et al, 2000;Somanathan et al, 2001;Sporbert et al, 2002;Solomon et al, 2004;Essers et al, 2005;Meister et al, 2007). The pattern of these foci changes with S-phase progression.…”

Section: Pcna Is Nuclear Throughout the Cell Cycle But Is Mainly Expmentioning

confidence: 99%

The distribution pattern of proliferating cell nuclear antigen in the nuclei of Leishmania donovani

et al. 2009

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“…Finally, we observed sensitivity to hydroxyurea (HU) in rad16-249 and rhp14Δ mutants, but again not in rad13Δ. HU causes fork stalling due to nucleotide depletion, and restart occurs via recombination-based mechanisms (Meister et al 2007;Lambert et al 2010;Sabatinos et al 2012).Slx4 and Saw1 are proposed to function as scaffolds that assemble XPF and other structure-specific endonucleases (Lyndaker and Alani 2009;Kashiyama et al 2013;Li et al 2013;Wan et al 2013). In contrast to budding yeast (Flott et al 2007;Li et al 2008), neither slx4Δ nor saw1Δ is sensitive to UV or MMS in fission yeast (Figure S3 and (Coulon et al 2006).…”

mentioning

confidence: 94%

Increased Meiotic Crossovers and Reduced Genome Stability in Absence of Schizosaccharomyces pombe Rad16 (XPF)

Mastro¹,

Forsburg

2014

Genetics

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Schizosaccharomyces pombe Rad16 is the ortholog of the XPF structure-specific endonuclease, which is required for nucleotide excision repair and implicated in the single strand annealing mechanism of recombination. We show that Rad16 is important for proper completion of meiosis. In its absence, cells suffer reduced spore viability and abnormal chromosome segregation with evidence for fragmentation. Recombination between homologous chromosomes is increased, while recombination within sister chromatids is reduced, suggesting that Rad16 is not required for typical homolog crossovers but influences the balance of recombination between the homolog and the sister. In vegetative cells, rad16 mutants show evidence for genome instability. Similar phenotypes are associated with mutants affecting Rhp14 XPA but are independent of other nucleotide excision repair proteins such as Rad13 XPG . Thus, the XPF/XPA module of the nucleotide excision repair pathway is incorporated into multiple aspects of genome maintenance even in the absence of external DNA damage.T HE XPF/ERCC1 protein complex is one of several structurespecific endonucleases that function broadly as resolvases in the repair of damaged DNA (Schwartz and Heyer 2011). Rad16/Swi9 is the fission yeast ortholog of endonuclease XPF, which forms a complex with Swi10/Rad23 (ERCC1) (Carr et al. 1994). XPF has a conserved role in nucleotide excision repair (NER) to remove UV-induced lesions in the DNA (Camenisch et al. 2006;Gregg et al. 2011). In humans, mutation of XPF is associated with xeroderma pigmentosum (XP), which causes sun sensitivity and high rates of skin cancer, as well as premature aging and neurological disorders (Camenisch et al. 2006;Gregg et al. 2011). Recent studies suggest XPF mutations are also associated with Fanconi anemia (FA), which requires repair of interstrand crosslinks (ICLs) (Bogliolo et al. 2013;Kashiyama et al. 2013). The canonical model for NER suggests that upon recognition of helix-distorting lesions, the DNA is unwound to form a bubble around the damage. XPA (SpRhp14) loads XPF (SpRad16) and ERCC1 (SpSwi10); XPF cleaves at the 59 end of the bubble while XPG (SpRad13), another endonuclease, cleaves at the 39 end to remove the offending segment (Fagbemi et al. 2011;Schwartz and Heyer 2011). Thus, Schizosaccharomyces pombe rad16 mutants show a decrease in (6-4) photoproduct excision (McCready et al. 1993;Carr et al. 1994).Consistent with these clinical effects, XPF is implicated in multiple mechanisms of genome maintenance (Paques and Haber 1997;Gregg et al. 2011;Schwartz and Heyer 2011). XPF is required for single strand annealing (SSA), a form of double strand break (DSB) repair distinct from typical homologous recombination (HR) (Ma et al. 2003;Kass and Jasin 2010). This occurs when short regions of homology exposed by resection are able to pair, leaving nonhomologous 39 overhangs as substrates for XPF cleavage. In budding yeast, recruitment of ScRad1 XPF and ScRad10 ERCC1 in this pathway depends on interactions with other pro...

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“…In Xenopus, the density of origin firing depends on ATM-and ATR-signaling pathways (25,26). In fission yeast, loss of Cds1 (the Chk2 homolog) results in an intriguing overlap of early and late DNA replication patterns (27). In mammalian cells, Chk1 participates in the intra-S-phase checkpoint that regulates late origins (28), but, even in an unperturbed S-phase, it represses the firing of origins located nearby replication forks.…”

Section: Mcms and The Control Of ''Excess'' Origins Of Replication: Amentioning

confidence: 99%

Excess MCM proteins protect human cells from replicative stress by licensing backup origins of replication

Ibarra

Schwob

Méndez

2008

Proc. Natl. Acad. Sci. U.S.A.

446

465

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The six main minichromosome maintenance proteins (Mcm2-7), which presumably constitute the core of the replicative DNA helicase, are present in chromatin in large excess relative to the number of active replication forks. To evaluate the relevance of this apparent surplus of Mcm2-7 complexes in human cells, their levels were down-regulated by using RNA interference. Interestingly, cells continued to proliferate for several days after the acute (>90%) reduction of Mcm2-7 concentration. However, they became hypersensitive to DNA replication stress, accumulated DNA lesions, and eventually activated a checkpoint response that prevented mitotic division. When this checkpoint was abrogated by the addition of caffeine, cells quickly lost viability, and their karyotypes revealed striking chromosomal aberrations. Singlemolecule analyses revealed that cells with a reduced concentration of Mcm2-7 complexes display normal fork progression but have lost the potential to activate ''dormant'' origins that serve a backup function during DNA replication. Our data show that the chromatin-bound ''excess'' Mcm2-7 complexes play an important role in maintaining genomic integrity under conditions of replicative stress.DNA combing ͉ DNA replication ͉ origin licensing R apidly proliferating cells start to prepare for DNA replication several hours before the actual S-phase, with the assembly of prereplication complexes (pre-RCs) at origins in telophase and early G 1 . Pre-RC assembly, also referred to as ''origin licensing,'' consists in the recruitment of Mcm2-7 protein complexes by initiator proteins ORC, CDC6, and CDT1. ORC and CDC6 likely constitute a structural module with ATPase activity that opens and closes the ring-shaped MCM hexamer, facilitating its topological engagement with the DNA, whereas CDT1 cooperates in the loading reaction as a molecular chaperone (reviewed in refs. 1 and 2). Different lines of evidence indicate that Mcm2-7 constitute the core of the replicative DNA helicase in eukaryotic cells in association with CDC45 and the GINS complex (3, 4).The maximum number of origins available in the subsequent S-phase is predetermined at the licensing stage, because additional pre-RCs cannot be assembled later in the cell cycle because of the inhibitory activity of the S, G 2 and M-phase cyclin-dependent kinases. This regulation establishes a temporal alternation of origin licensing and firing that is important to prevent DNA overreplication. In yeast, blending the licensing and firing periods by deletion of the CDK inhibitor Sic1 or by overexpression of the G 1 cyclin Cln2, greatly increased genomic instability (5, 6). In human cells, premature expression of Cyclin E during mitosis and G 1 interfered with the association of MCM proteins with chromatin and at the same time promoted the firing of the limited number of licensed origins, effectively accelerating the G 1 -S transition (7). Nevertheless, cells continued to proliferate under these challenging conditions and accumulated karyotypic defects (8). These results are ...

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Replication foci dynamics: replication patterns are modulated by S-phase checkpoint kinases in fission yeast

Cited by 78 publications

References 47 publications

The distribution pattern of proliferating cell nuclear antigen in the nuclei of Leishmania donovani

The distribution pattern of proliferating cell nuclear antigen in the nuclei of Leishmania donovani

Increased Meiotic Crossovers and Reduced Genome Stability in Absence of Schizosaccharomyces pombe Rad16 (XPF)

Excess MCM proteins protect human cells from replicative stress by licensing backup origins of replication

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