Human single-stranded DNA binding protein 1 (hSSB1/NABP2) is required for the stability and repair of stalled replication forks

Aberrant DNA replication is a primary cause of mutations that are associated with pathological disorders including cancer. During DNA metabolism, the primary causes of replication fork stalling include secondary DNA structures, highly transcribed regions and damaged DNA. The restart of stalled replication forks is critical for the timely progression of the cell cycle and ultimately for the maintenance of genomic stability. Our previous work has implicated the single-stranded DNA binding protein, hSSB1/NABP2, in the repair of DNA double-strand breaks via homologous recombination. Here, we demonstrate that hSSB1 relocates to hydroxyurea (HU)-damaged replication forks where it is required for ATR and Chk1 activation and recruitment of Mre11 and Rad51. Consequently, hSSB1-depleted cells fail to repair and restart stalled replication forks. hSSB1 deficiency causes accumulation of DNA strand breaks and results in chromosome aberrations observed in mitosis, ultimately resulting in hSSB1 being required for survival to HU and camptothecin. Overall, our findings demonstrate the importance of hSSB1 in maintaining and repairing DNA replication forks and for overall genomic stability

Role of replication protein A as sensor in activation of the S-phase checkpoint in Xenopus egg extracts

Uncoupling between DNA polymerases and helicase activities at replication forks, induced by diverse DNA lesions or replication inhibitors, generate long stretches of primed single-stranded DNA that is implicated in activation of the S-phase checkpoint. It is currently unclear whether nucleation of the essential replication factor RPA onto this substrate stimulates the ATR-dependent checkpoint response independently of its role in DNA synthesis. Using Xenopus egg extracts to investigate the role of RPA recruitment at uncoupled forks in checkpoint activation we have surprisingly found that in conditions in which DNA synthesis occurs, RPA accumulation at forks stalled by either replication stress or UV irradiation is dispensable for Chk1 phosphorylation. In contrast, when both replication fork uncoupling and RPA hyperloading are suppressed, Chk1 phosphorylation is inhibited. Moreover, we show that extracts containing reduced levels of RPA accumulate ssDNA and induce spontaneous, caffeine-sensitive, Chk1 phosphorylation in S-phase. These results strongly suggest that disturbance of enzymatic activities of replication forks, rather than RPA hyperloading at stalled forks, is a critical determinant of ATR activation

Solidaires; essai de sociologie chrétienne.

Mode of access: Internet

Implication of RPA32 phosphorylation in S-phase checkpoint signalling at replication forks stalled with aphidicolin in Xenopus egg extracts

International audienc

Des vestiges d’habitat au sud de la voie principale

International audienc

Les thermes publics

International audienc

Médicaments contrefaits (la France épargnée ?)

BORDEAUX2-BU Santé (330632101) / SudocSudocFranceF

Soudage par Faisceau d’Electrons de l’Acier Inoxydable Austénitique stabilisé au niobium X6CrNiMoNb 17.12.2

DCNS a conduit une démarche complète de conception et de réalisation d’un équipement sous pression nucléaire en acier inoxydable austénitique stabilisé au niobium (Nb) en remplacement d’une technologie acier au carbone revêtu d’acier inoxydable. Ce changement de technologie a conduit en particulier à des travaux portant sur la qualification du soudage des soudures principales par faisceau d’électrons (FE) en forte épaisseur, et sur la contrôlabilité de ces soudures en fabrication et en service. La présentation développera la démarche de qualification du mode opératoire de soudage et présentera les résultats obtenus pour les soudures de production

Soudage par Faisceau d’Electrons de l’Acier Inoxydable Austenitique stabilise au niobium X6CrNiMoNb 17.12.2

DCNS a conduit une démarche complète de conception et de réalisation d’un équipement sous pression nucléaire en acier inoxydable austénitique stabilisé au niobium (Nb) en remplacement d’une technologie acier au carbone revêtu d’acier inoxydable. Ce changement de technologie a conduit en particulier à des travaux portant sur la qualification du soudage des soudures principales par faisceau d’électrons (FE) en forte épaisseur, et sur la contrôlabilité de ces soudures en fabrication et en service