Dampening
            <scp>DNA</scp>
            damage checkpoint signalling via coordinated
            <scp>BRCT</scp>
            domain interactions

Cussiol, José Renato Rosa; Jablonowski, Carolyn M; Yimit, Askar; Brown, Grant W.; Smolka, Marcus B.

doi:10.15252/embj.201490834

Cited by 46 publications

(76 citation statements)

References 55 publications

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“…Cells were then treated with MMS for 2 hr and then centrifuged and recovered in fresh, MMS-free medium for up to 6 hr. For a detailed protocol please see Cussiol et al (2015).…”

Section: Pulsed-field Gel Electrophoresismentioning

confidence: 99%

“…Later on, the PP4 phosphatase complex Pph3-Psy2 was shown to be important for Rad53 dephosphorylation following treatment with the DNA alkylating agent methyl methanesulfonate (MMS), which generates replication blocks that are readily bypassed by moving replication forks (O'Neill et al 2007). In addition to phosphatase-mediated mechanisms, we have recently uncovered a new mechanism of Rad53 downregulation involving direct displacement of Rad9 from DNA lesions (Ohouo et al 2013;Cussiol et al 2015). In this phosphatase-independent mechanism, named dampens adaptormediated phosphosignaling (DAMP), a complex formed by the DNA repair scaffolds Slx4 and Rtt107 competes with Rad9 by interacting with two proteins required for Rad9 recruitment, namely g-H2A and Dpb11.…”

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confidence: 99%

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Termination of Replication Stress Signaling via Concerted Action of the Slx4 Scaffold and the PP4 Phosphatase

et al. 2015

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In response to replication stress, signaling mediated by DNA damage checkpoint kinases protects genome integrity. However, following repair or bypass of DNA lesions, checkpoint signaling needs to be terminated for continued cell cycle progression and proliferation. In budding yeast, the PP4 phosphatase has been shown to play a key role in preventing hyperactivation of the checkpoint kinase Rad53. In addition, we recently uncovered a phosphatase-independent mechanism for downregulating Rad53 in which the DNA repair scaffold Slx4 decreases engagement of the checkpoint adaptor Rad9 at DNA lesions. Here we reveal that proper termination of checkpoint signaling following the bypass of replication blocks imposed by alkylated DNA adducts requires the concerted action of these two fundamentally distinct mechanisms of checkpoint downregulation. Cells lacking both SLX4 and the PP4-subunit PPH3 display a synergistic increase in Rad53 signaling and are exquisitely sensitive to the DNA alkylating agent methyl methanesulfonate, which induces replication blocks and extensive formation of chromosomal linkages due to template switching mechanisms required for fork bypass. Rad53 hypersignaling in these cells seems to converge to a strong repression of Mus81-Mms4, the endonuclease complex responsible for resolving chromosomal linkages, thus explaining the selective sensitivity of slx4D pph3D cells to alkylation damage. Our results support a model in which Slx4 acts locally to downregulate Rad53 activation following fork bypass, while PP4 acts on pools of active Rad53 that have diffused from the site of lesions. We propose that the proper spatial coordination of the Slx4 scaffold and PP4 action is crucial to allow timely activation of Mus81-Mms4 and, therefore, proper chromosome segregation.

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“…Cells were then treated with MMS for 2 hr and then centrifuged and recovered in fresh, MMS-free medium for up to 6 hr. For a detailed protocol please see Cussiol et al (2015).…”

Section: Pulsed-field Gel Electrophoresismentioning

confidence: 99%

mentioning

confidence: 99%

Termination of Replication Stress Signaling via Concerted Action of the Slx4 Scaffold and the PP4 Phosphatase

et al. 2015

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“…77,79 It appears that BRCT1-2 and BRCT3-4 of Dpb11 can engage Slx4 and the 9-1-1 complex. 77,79,80 As such, Dpb11 can bridge Slx4 and 9-1-1 (Fig. 3).…”

Section: The Rtt107 and Slx4 Association And Its Functions In Conjuncmentioning

confidence: 97%

“…3). 77,80,81 An underlying mechanism may be that Slx4 and Rad9 compete for binding to BRCT1-2 of Dpb11, though Slx4 can also associate with BRCT3-4 of Dpb11. 77,79 It appears that BRCT1-2 and BRCT3-4 of Dpb11 can engage Slx4 and the 9-1-1 complex.…”

Section: The Rtt107 and Slx4 Association And Its Functions In Conjuncmentioning

confidence: 99%

“…3). 80,82 It is possible that the affinity of the tBRCT of Rad9 toward gH2A is less than that of Rtt107, a hypothesis worthy of additional testing. Despite this potential difference, BRCT5-6 of Rtt107 and tBRCT of Rad9 are interchangeable and the domain swapped constructs can partially support the functions of either protein.…”

Section: The Rtt107 and Slx4 Association And Its Functions In Conjuncmentioning

confidence: 99%

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Multi-BRCT scaffolds use distinct strategies to support genome maintenance

2016

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Genome maintenance requires coordinated actions of diverse DNA metabolism processes. Scaffolding proteins, such as those containing multiple BRCT domains, can influence these processes by collaborating with numerous partners. The best-studied examples of multi-BRCT scaffolds are the budding yeast Dpb11 and its homologues in other organisms, which regulate DNA replication, repair, and damage checkpoints. Recent studies have shed light on another group of multi-BRCT scaffolds, including Rtt107 in budding yeast and related proteins in other organisms. These proteins also influence several DNA metabolism pathways, though they use strategies unlike those employed by the Dpb11 family of proteins. Yet, at the same time, these 2 classes of multi-BRCT proteins can collaborate under specific situations. This review summarizes recent advances in our understanding of how these multi-BRCT proteins function in distinct manners and how they collaborate, with a focus on Dpb11 and Rtt107.

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A field guide to the proteomics of post‐translational modifications in DNA repair

Sanford

Smolka

2022

Proteomics

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All cells incur DNA damage from exogenous and endogenous sources and possess pathways to detect and repair DNA damage. Post-translational modifications (PTMs), in the past 20 years, have risen to ineluctable importance in the study of the regulation of DNA repair mechanisms. For example, DNA damage response kinases are critical in both the initial sensing of DNA damage as well as in orchestrating downstream activities of DNA repair factors. Mass spectrometry-based proteomics revolutionized the study of the role of PTMs in the DNA damage response and has canonized PTMs as central modulators of nearly all aspects of DNA damage signaling and repair. This review provides a biologist-friendly guide for the mass spectrometry analysis of PTMs in the context of DNA repair and DNA damage responses. We reflect on the current state of proteomics for exploring new mechanisms of PTM-based regulation and outline a roadmap for designing PTM mapping experiments that focus on the DNA repair and DNA damage responses.

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Dampening DNA damage checkpoint signalling via coordinated BRCT domain interactions

Cited by 46 publications

References 55 publications

Termination of Replication Stress Signaling via Concerted Action of the Slx4 Scaffold and the PP4 Phosphatase

Termination of Replication Stress Signaling via Concerted Action of the Slx4 Scaffold and the PP4 Phosphatase

Multi-BRCT scaffolds use distinct strategies to support genome maintenance

A field guide to the proteomics of post‐translational modifications in DNA repair

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