TUdatalib : The human Timeless-Tipin complex assures the replication fork protection ahead of the replisome

Zur Kurzanzeige

dc.contributor.author	Prorok, Paulina
dc.contributor.author	Wolf, Eva
dc.contributor.author	Cardoso, M. Cristina
dc.date.accessioned	2023-11-29T12:58:55Z
dc.date.available	2023-11-29T12:58:55Z
dc.date.issued	2023
dc.identifier.uri	https://tudatalib.ulb.tu-darmstadt.de/handle/tudatalib/4026
dc.identifier.uri	https://doi.org/10.48328/tudatalib-1282
dc.description	The accuracy of replication is one of the most important mechanisms ensuring stability of the genome. The fork protection complex prevents premature replisome stalling and/or premature disassembly upon stress. Here, we characterize the Timeless-Tipin complex, a recently identified fork protection component. We used microscopy approaches including colocalization analysis and proximity ligation assay to investigate the spatial localization of the complex during ongoing replication in human cells. Taking advantage of the replication stress induction and the ensuing polymerase-helicase decoupling, we characterized the Timeless-Tipin localization within the replisome. Replication stress was induced using hydroxyurea (HU) and aphidicolin (APH). While HU depletes the substrate for DNA synthesis, APH binds directly inside the catalytic pocket of DNA polymerase and inhibits its activity. Our data revealed that the Timeless-Tipin complex, independently of the stress, remains bound on chromatin upon stress induction and progresses together with the replicative helicase. This is accompanied by the spatial dissociation of the complex from the blocked replication machinery. Additionally, after stress induction, Timeless interaction with RPA, which continuously accumulates on ssDNA, was increased. Taken together, the Timeless-Tipin complex acts as an universal guardian of the mammalian replisome, in an unperturbed S phase progression as well as during replication stress.	de_DE
dc.language.iso	en	de_DE
dc.rights	Creative Commons Attribution-NonCommercial 4.0
dc.rights.uri	https://creativecommons.org/licenses/by-nc/4.0/
dc.subject	Timeless-Tipin complex	de_DE
dc.subject	fork protection complex	de_DE
dc.subject	DNA replication stress	de_DE
dc.subject	helicase-polymerase decoupling	de_DE
dc.subject	aphidicolin	de_DE
dc.subject	hydroxyurea	de_DE
dc.subject	proximity ligation assay	de_DE
dc.subject.classification	2.11-03 Zellbiologie	de_DE
dc.subject.ddc	570
dc.title	The human Timeless-Tipin complex assures the replication fork protection ahead of the replisome	de_DE
dc.type	Image	de_DE
dc.description.version	1st submission	de_DE
tud.unit	TUDa
tud.history.classification	Version=2020-2024;201-03 Zellbiologie