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Università della Svizzera italiana

Crossover or non-crossover outcomes : tailored processing of homologous recombination intermediates

Sanchez, Aurore ; Reginato, Giordano ; Cejka, Petr

In: Current opinion in genetics & development, 2021, vol. 71, p. 39-47

DNA breaks may arise accidentally in vegetative cells or in a programmed manner in meiosis. The usage of a DNA template makes homologous recombination potentially error-free, however, recombination is not always accurate. Cells possess a remarkable capacity to tailor processing of recombination intermediates to fulfill a particular need. Vegetatively growing cells aim to maintain genome...

Università della Svizzera italiana

Main steps in DNA double-strand break repair : An introduction to homologous recombination and related processes

Ranjha, Lepakshi ; Howard, Sean Michael ; Cejka, Petr

In: Chromosoma, 2018, vol. 127, no. 2 (June), p. 187–214

DNA double-strand breaks arise accidentally upon exposure of DNA to radiation, chemicals or result from faulty DNA metabolic processes. DNA breaks can also be introduced in a programmed manner, such as during the maturation of the immune system, meiosis or cancer chemo- or radiotherapy. Cells have developed a variety of repair pathways, which are fine-tuned to the specific needs of a cell....

Università della Svizzera italiana

A meiotic XPF-ERCC1-like complex recognizes joint molecule recombination intermediates to promote crossover formation

De Muyt, Arnaud ; Pyatnitskaya, Alexandra ; Andréani, Jessica ; Ranjha, Lepakshi ; Laureau, Raphaëlle ; Fernandez-Vega, Ambra ; Holoch, Daniel ; Girard, Elodie ; Govin, Jérome ; Margueron, Raphaël ; Couté, Yohann ; Cejka, Petr ; Guérois, Raphaël ; Borde, Valérie

In: Genes and development, 2018, vol. 32, no. 3-4, p. 283-296

Meiotic crossover formation requires the stabilization of early recombination intermediates by a set of proteins and occurs within the environment of the chromosome axis, a structure important for the regulation of meiotic recombination events. The molecular mechanisms underlying and connecting crossover recombination and axis localization are elusive. Here, we identified the ZZS...

Università della Svizzera italiana

The motor activity of DNA2 functions as an ssDNA translocase to promote DNA end resection

Levikova, Maryna ; Pinto, Cosimo ; Cejka, Petr

In: Genes and Development, 2017, vol. 31, no. 5, p. 493-502

DNA2 nuclease–helicase functions in DNA replication and recombination. This requires the nuclease of DNA2, while, in contrast, the role of the helicase activity has been unclear. We now show that the motor activity of both recombinant yeast and human DNA2 promotes efficient degradation of long stretches of ssDNA, particularly in the presence of the replication protein A. This degradation is...

Università della Svizzera italiana

The MRE11 complex : a versatile toolkit for the repair of broken DNA

Reginato, Giordano ; Cejka, Petr

In: DNA repair, 2020, vol. 91–92, no. July–August, p. 13 p

When DNA breaks, the ends need to be stabilized and processed to facilitate subsequent repair, which can occur by either direct but error-prone end-joining with another broken DNA molecule or a more accurate homology-directed repair by the recombination machinery. At the same time, the presence of broken DNA triggers a signaling cascade that regulates the repair events and cellular progression...

Università della Svizzera italiana

The Mre11-Nbs1 interface is essential for viability and tumor suppression

Kim, Jun Hyun ; Grosbart, Malgorzata ; Anand, Roopesh ; Wyman, Claire ; Cejka, Petr ; Petrini, John H.J.

In: Cell reports, 2017, vol. 18, no. 2, p. 496-507

The Mre11 complex (Mre11, Rad50, and Nbs1) is integral to both DNA repair and ataxia telangiectasia mutated (ATM)-dependent DNA damage signaling. All three Mre11 complex components are essential for viability at the cellular and organismal levels. To delineate essential and non-essential Mre11 complex functions that are mediated by Nbs1, we used TALEN-based genome editing to derive Nbs1...

Università della Svizzera italiana

NBS1 promotes the endonuclease of the MRE11-RAD50 complex by sensing CtIP phosphorylation

Anand, Roopesh ; Jasrotia, Arti ; Bundschuh, Diana ; Howard, Sean Michael ; Ranjha, Lepakshi ; Stucki, Manuel ; Cejka, Petr

In: The EMBO Journal, 2019, vol. 38, no. 7, p. e101005

DNA end resection initiates DNA break repair by homologous recombination. MRE11-RAD50-NBS1 and phosphorylated CtIP perform the first resection step by MRE11-catalyzed endonucleolytic DNA cleavage. Human NBS1, more than its Xrs2 homologue from Saccharomyces cerevisiae, is crucial for this process, highlighting complex mechanisms that regulate the MRE11 nuclease in high eukaryotes. Using a...

Università della Svizzera italiana

Physiological protein blocks direct the Mre11–Rad50–Xrs2 and Sae2 nuclease complex to initiate DNA end resection

Reginato, Giordano ; Cannavo, Elda ; Cejka, Petr

In: Genes and Development, 2017, vol. 31, no. 23-24, p. 2325-2330

DNA double-strand break repair by homologous recombination is initiated by DNA end resection, which is commenced by the Mre11–Rad50–Xrs2 complex and Sae2 in yeast. Here we report that the nonhomologous end joining factor Ku limits the exonuclease activity of Mre11 and promotes its endonuclease to cleave 5'-terminated DNA strands at break sites. Following initial endonucleolytic cleavage...

Università della Svizzera italiana

Regulatory control of DNA end resection by Sae2 phosphorylation

Cannavo, Elda ; Johnson, Dominic ; Andres, Sara N. ; Kissling, Vera M. ; Reinert, Julia K. ; Garcia, Valerie ; Erie, Dorothy A. ; Hess, Daniel ; Thomä, Nicolas H. ; Enchev, Radoslav I. ; Peter, Matthias ; Williams, R. Scott ; Neale, Matt J. ; Cejka, Petr

In: Nature communications, 2018, vol. 9, p. 4016

DNA end resection plays a critical function in DNA double-strand break repair pathway choice. Resected DNA ends are refractory to end-joining mechanisms and are instead channeled to homology-directed repair. Using biochemical, genetic, and imaging methods, we show that phosphorylation of Saccharomyces cerevisiae Sae2 controls its capacity to promote the Mre11-Rad50-Xrs2 (MRX) nuclease to...

Università della Svizzera italiana

Restoration of replication fork stability in BRCA1- and BRCA2-deficient cells by inactivation of SNF2-family fork remodelers

Taglialatela, Angelo ; Alvarez, Silvia ; Leuzzi, Giuseppe ; Sannino, Vincenzo ; Ranjha, Lepakshi ; Huang, Jen-Wei ; Madubata, Chioma ; Anand, Roopesh ; Levy, Brynn ; Rabadan, Raul ; Cejka, Petr ; Costanzo, Vincenzo ; Ciccia, Alberto

In: Molecular Cell, 2017, vol. 68, no. 2, p. 414-430.e8

To ensure the completion of DNA replication and maintenance of genome integrity, DNA repair factors protect stalled replication forks upon replication stress. Previous studies have identified a critical role for the tumor suppressors BRCA1 and BRCA2 in preventing the degradation of nascent DNA by the MRE11 nuclease after replication stress. Here we show that depletion of SMARCAL1, a...