MTBP phosphorylation controls DNA replication origin firing

Ferreira, Pedro; Höfer, Verena; Kronshage, Nora; Marko, Anika; Reusswig, Karl-Uwe; Tetik, Bilal; Dießel, Christoph; Köhler, Kerstin; Tschernoster, Nikolai; Altmüller, Janine; Schulze, Nina; Pfander, Boris; Boos, Dominik

doi:10.1038/s41598-021-83287-w

Cited by 12 publications

(21 citation statements)

References 82 publications

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“…Inhibiting ATR/ATM alone had a smaller effect on MTBP dephosphorylation whereas Chk1 inhibition alone had no visible effect on MTBP phosphorylation ( Supplementary Figure S7 ). This observation could be explained by a cooperative effect of the checkpoint kinases as concluded in a recent study, published while our work was in revision ( 55 ). In yeast, the Treslin homologue Sld3 shifts in a DDK dependent manner in G1 phase ( 56 ).…”

Section: Resultssupporting

confidence: 70%

Polo-like kinase 1 (Plk1) regulates DNA replication origin firing and interacts with Rif1 in Xenopus

Ciardo

Haccard

Narassimprakash

et al. 2021

Nucleic Acids Research

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The activation of eukaryotic DNA replication origins needs to be strictly controlled at multiple steps in order to faithfully duplicate the genome and to maintain its stability. How the checkpoint recovery and adaptation protein Polo-like kinase 1 (Plk1) regulates the firing of replication origins during non-challenged S phase remained an open question. Using DNA fiber analysis, we show that immunodepletion of Plk1 in the Xenopus in vitro system decreases replication fork density and initiation frequency. Numerical analyses suggest that Plk1 reduces the overall probability and synchrony of origin firing. We used quantitative chromatin proteomics and co-immunoprecipitations to demonstrate that Plk1 interacts with firing factors MTBP/Treslin/TopBP1 as well as with Rif1, a known regulator of replication timing. Phosphopeptide analysis by LC/MS/MS shows that the C-terminal domain of Rif1, which is necessary for its repressive action on origins through protein phosphatase 1 (PP1), can be phosphorylated in vitro by Plk1 on S2058 in its PP1 binding site. The phosphomimetic S2058D mutant interrupts the Rif1-PP1 interaction and modulates DNA replication. Collectively, our study provides molecular insights into how Plk1 regulates the spatio-temporal replication program and suggests that Plk1 controls origin activation at the level of large chromatin domains in vertebrates.

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Section: Resultssupporting

confidence: 70%

Polo-like kinase 1 (Plk1) regulates DNA replication origin firing and interacts with Rif1 in Xenopus

Ciardo

Haccard

Narassimprakash

et al. 2021

Nucleic Acids Research

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“…Quantification of replicates (Fig. 2C) confirmed these observations and proved reproducibility of our BrdU quantification method (Boos et al 2013, Ferreira et al 2021, Kohler et al 2019. Treslin/TICRR-DSTD clone 21 rescued replication somewhat better (50 % replication) than clones 11 and 17 (approximately 30 % replication; 2PM and no-transgene controls about 30%), exemplifying our observation that individual clones expressing the same transgene showed some variability that probably arise through clonal selection.…”

Section: Resultssupporting

confidence: 78%

Refining the domain architecture model of the replication origin firing factor Treslin/TICRR

Ferreira

Sánchez‐Pulido

Marko

et al. 2021

Preprint

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Faithful genome duplication requires appropriately controlled replication origin firing. The metazoan Treslin/TICRR origin firing factor and its yeast orthologue Sld3 are regulation hubs of origin firing. They share the Sld3-Treslin domain (STD) and the adjacent TopBP1/Dpb11 interaction domain (TDIN). We report a revised domain architecture model of Treslin/TICRR. Complementary protein sequence analyses uncovered Ku70-homologous lower case Greek beta-barrel folds in the Treslin/TICRR middle domain (M domain) and in Sld3. Thus, the Sld3-homologous Treslin/TICRR core comprises its three central domains, M domain, STD and TDIN. This Sld3-core is flanked by non-conserved terminal domains, the CIT (conserved in Treslins) and the C-terminus. We also identified Ku70-like lower case Greek beta-barrels in MTBP and Sld7. Our binding experiments showed that the Treslin lower case Greek beta-barrel mediates interaction with the MTBP lower case Greek beta-barrel, reminiscent of the homotypic Ku70-Ku80 dimerization. This binding mode is conserved in the Sld3-Sld7 dimer. We used Treslin/TICRR domain mutants to show that all Sld3-core domains and the non-conserved terminal domains fulfil important functions during origin firing in human cells. Thus, metazoa-specific and widely conserved molecular processes cooperate during origin firing in metazoa.

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“…In contrast, the requirement of the Sld3-homologous STD for replication had not previously been addressed in higher eukaryotes. To test this, we used incorporation of the nucleotide analogue BrdU into nascent DNA of cultured human cells in an established RNAireplacement system (Boos et al, 2011(Boos et al, , 2013 (Boos et al, 2013;K öhler et al, 2019;Ferreira et al, 2021) confirmed these observations. Treslin/TICRR-ΔSTD clone 21 rescued replication somewhat better (50% replication) than clones 11 (~30% (Boos et al, 2011;Kumagai et al, 2011), Chk1 binds to the very C-terminal 99 amino acids of Treslin (Guo et al, 2015), and Brd2/4 binds to the Treslin/TICRR region 1560-1580 (Sansam et al, 2018).…”

Section: Resultsmentioning

confidence: 93%

Refining the domain architecture model of the replication origin firing factor Treslin/TICRR

et al. 2022

Self Cite

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Faithful genome duplication requires appropriately controlled replication origin firing. The metazoan origin firing regulation hub Treslin/TICRR and its yeast orthologue Sld3 share the Sld3-Treslin domain and the adjacent TopBP1/Dpb11 interaction domain. We report a revised domain architecture model of Treslin/TICRR. Protein sequence analyses uncovered a conserved Ku70-homologous β-barrel fold in the Treslin/TICRR middle domain (M domain) and in Sld3. Thus, the Sld3-homologous Treslin/TICRR core comprises its three central domains, M domain, Sld3-Treslin domain, and TopBP1/Dpb11 interaction domain, flanked by non-conserved terminal domains, the CIT (conserved in Treslins) and the C terminus. The CIT includes a von Willebrand factor type A domain. Unexpectedly, MTBP, Treslin/TICRR, and Ku70/80 share the same N-terminal domain architecture, von Willebrand factor type A and Ku70-like β-barrels, suggesting a common ancestry. Binding experiments using mutants and the Sld3–Sld7 dimer structure suggest that the Treslin/Sld3 and MTBP/Sld7 β-barrels engage in homotypic interactions, reminiscent of Ku70-Ku80 dimerization. Cells expressing Treslin/TICRR domain mutants indicate that all Sld3-core domains and the non-conserved terminal domains fulfil important functions during origin firing in human cells. Thus, metazoa-specific and widely conserved molecular processes cooperate during metazoan origin firing.

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MTBP phosphorylation controls DNA replication origin firing

Cited by 12 publications

References 82 publications

Polo-like kinase 1 (Plk1) regulates DNA replication origin firing and interacts with Rif1 in Xenopus

Polo-like kinase 1 (Plk1) regulates DNA replication origin firing and interacts with Rif1 in Xenopus

Refining the domain architecture model of the replication origin firing factor Treslin/TICRR

Refining the domain architecture model of the replication origin firing factor Treslin/TICRR

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