Cited by 240DNA Repair MechanismsCancer-related Molecular PathwaysGenomics and Chromatin Dynamics

The Human Tim/Tipin Complex Coordinates an Intra-S Checkpoint Response to UV That Slows Replication Fork Displacement

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Ünsal-Kaçmaz, Keziban, Chastain, Paul D., Qu, Ping-Ping, Minoo, Parviz, Cordeiro-Stone, Marila, Sancar, Aziz, Kaufmann, William K.·2007·Molecular and Cellular Biology

Abstract

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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  1. 22a.Heffernan, T. P., K. Ünsal-Kaçmaz, D. A. Simpson, A. N. Heinloth, R. S. Paules, A. Sancar, M. Cordeiro-Stone, and W. K. Kaufmann. J. Biol. Chem., in press. [DOI] [PMC free article] [PubMed]

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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
Published
2007-04-01
Volume
27
Issue
8
Pages
3131-3142
ISSN
  • 0270-7306 (ISSN-L)
  • 1098-5549
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
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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