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The Human Tim/Tipin Complex Coordinates an Intra-S Checkpoint Response to UV That Slows Replication Fork Displacement
Open accessAbstract
UV-induced DNA damage stalls DNA replication forks and activates the intra-S checkpoint to inhibit replicon initiation. In response to stalled replication forks, ATR phosphorylates and activates the transducer kinase Chk1 through interactions with the mediator proteins TopBP1, Claspin, and Timeless (Tim). Murine Tim recently was shown to form a complex with Tim-interacting protein (Tipin), and a similar complex was shown to exist in human cells. Knockdown of Tipin using small interfering RNA reduced the expression of Tim and reversed the intra-S checkpoint response to UVC. Tipin interacted with replication protein A (RPA) and RPA-coated DNA, and RPA promoted the loading of Tipin onto RPA-free DNA. Immunofluorescence analysis of spread DNA fibers showed that treating HeLa cells with 2.5 J/m(2) UVC not only inhibited the initiation of new replicons but also reduced the rate of chain elongation at active replication forks. The depletion of Tim and Tipin reversed the UV-induced inhibition of replicon initiation but affected the rate of DNA synthesis at replication forks in different ways. In undamaged cells depleted of Tim, the apparent rate of replication fork progression was 52% of the control. In contrast, Tipin depletion had little or no effect on fork progression in unirradiated cells but significantly attenuated the UV-induced inhibition of DNA chain elongation. Together, these findings indicate that the Tim-Tipin complex mediates the UV-induced intra-S checkpoint, Tim is needed to maintain DNA replication fork movement in the absence of damage, Tipin interacts with RPA on DNA and, in UV-damaged cells, Tipin slows DNA chain elongation in active replicons.
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“The circadian protein Tim (Timeless) -Tipin (Timeless-interacting protein) complex associates with ATR-Chk1 checkpoint proteins and plays an essential role in DDR as well [130–133].”
From Functional interplay between ATM/ATR-mediated DNA damage response and DNA repair pathways in oxidative stress (2014) · cites this paper as [131]
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Metadata
- Authors
- Ünsal-Kaçmaz, K., Chastain, P. D., Qu, P.-P., Minoo, P., Cordeiro-Stone, M., Sancar, A., & Kaufmann, W. K.
- Affiliations
- University of North Carolina at Chapel Hill (US)
- University of Southern California (US)
- Department of Pathology and Laboratory Medicine, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599
- Corresponding author
- Kaufmann, William K.
- Venue
- Molecular and Cellular Biology
- Publisher
- Informa UK Limited
- Published
- 2007-04-01
- Volume
- 27
- Issue
- 8
- Pages
- 3131-3142
- ISSN
- 0270-7306 (ISSN-L)
- 1098-5549
- PMC
- PMC1899931
- OpenAlex
- W2127622729
- Cited by
- 240
- References
- 67
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 9 · 2023: 9 · 2022: 6 · 2021: 6 · 2020: 7 · 2019: 10 · 2018: 3 · 2017: 10 · 2016: 15 · 2015: 13 · 2014: 23 · 2013: 19 · 2012: 13
- FWCI
- 10.49
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Molecular and Cellular Biology (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Carolina Digital Repository (University of North Carolina at Chapel Hill) (submittedVersion)
- UNC Libraries
- Funders
- National Cancer Institute (R01CA055065, CA 55065, CA 81343, R01CA081343, P30CA016086)
- National Institute of General Medical Sciences (R01GM032833, GM32833)
- National Institute of Environmental Health Sciences (R01ES011012, ES11012, P30ES010126)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › DNA Repair Mechanisms
- Health Sciences › Medicine › Oncology › Cancer-related Molecular Pathways
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
- Keywords
- S-phase checkpoint, UV-induced DNA damage, replication fork stalling, replication fork progression, DNA chain elongation, ATR-Chk1 signaling, TopBP1, Claspin, TIMELESS, replication protein A, UV-C radiation, DNA damage response
- MeSH terms
- Checkpoint Kinase 1, Amino Acid Sequence, Carrier Proteins, DNA, DNA Replication, DNA-Binding Proteins, Enzyme Activation, HeLa Cells, Humans, Idoxuridine, Models, Genetic, Molecular Sequence Data, Nuclear Proteins, Protein Binding, Protein Kinases, Ultraviolet Rays, Down-Regulation, S Phase, Cell Cycle Proteins, Intracellular Signaling Peptides and Proteins, Replication Protein A, Xeroderma Pigmentosum Group A Protein
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