Transcription-Associated Recombination Is Dependent on Replication in Mammalian Cells

ABSTRACT Transcription can enhance recombination; this is a ubiquitous phenomenon from prokaryotes to higher eukaryotes. However, the mechanism of transcription-associated recombination in mammalian cells is poorly understood. Here we have developed a construct with a recombination substrate in which levels of recombination can be studied in the presence or absence of transcription. We observed a direct enhancement in recombination when transcription levels through the substrate were increased. This increase in homologous recombination following transcription is locus specific, since homologous recombination at the unrelated hprt gene is unaffected. In addition, we have shown that transcription-associated recombination involves both short-tract and long-tract gene conversions in mammalian cells, which are different from double-strand-break-induced recombination events caused by endonucleases. Transcription fails to enhance recombination in cells that are not in the S phase of the cell cycle. Furthermore, inhibition of transcription suppresses induction of recombination at stalled replication forks, suggesting that recombination may be involved in bypassing transcription during replication.

Download Full-text

158 Double strand break repair in mammalian cells: competition between homologous recombination and non-homologous end joining through cell cycle

Radiotherapy and Oncology ◽

10.1016/s0167-8140(04)82025-5 ◽

2004 ◽

Vol 73 ◽

pp. S73-S74

Keyword(s):

Cell Cycle ◽

Homologous Recombination ◽

Mammalian Cells ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

End Joining ◽

Break Repair ◽

Non Homologous End Joining

Download Full-text

Faculty Opinions recommendation of Ctp1 is a cell-cycle-regulated protein that functions with Mre11 complex to control double-strand break repair by homologous recombination.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.1093906.548940 ◽

2007 ◽

Author(s):

Lucio Comai

Keyword(s):

Cell Cycle ◽

Homologous Recombination ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

Mre11 Complex ◽

A Cell ◽

Break Repair

Download Full-text

Double strand break repair by homologous recombination is regulated by cell cycle-independent signaling via ATM in human glioma cells. Vol. 279 (2004) 15402-15410

Journal of Biological Chemistry ◽

10.1016/s0021-9258(20)66633-9 ◽

2004 ◽

Vol 279 (23) ◽

pp. 24906

Author(s):

Sarah E. Golding ◽

Elizabeth Rosenberg ◽

Ashraf Khalil ◽

Alison McEwen ◽

Matthew Holmes ◽

...

Keyword(s):

Cell Cycle ◽

Homologous Recombination ◽

Glioma Cells ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

Human Glioma ◽

Human Glioma Cells ◽

Break Repair

Download Full-text

Double-Strand Break Repair and Homologous Recombination in Mammalian Cells

DNA Damage and Repair ◽

10.1385/1-59259-095-0:207 ◽

2003 ◽

pp. 207-236 ◽

Cited By ~ 7

Author(s):

Maria Jasin

Keyword(s):

Homologous Recombination ◽

Mammalian Cells ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

Break Repair

Download Full-text

Evidence for DNA-PK-dependent and -independent DNA double-strand break repair pathways in mammalian cells as a function of the cell cycle.

Molecular and Cellular Biology ◽

10.1128/mcb.17.3.1425 ◽

1997 ◽

Vol 17 (3) ◽

pp. 1425-1433 ◽

Cited By ~ 146

Author(s):

S E Lee ◽

R A Mitchell ◽

A Cheng ◽

E A Hendrickson

Keyword(s):

Cell Cycle ◽

Mammalian Cells ◽

Cellular Response ◽

Strand Break ◽

S Phase ◽

Dependent Manner ◽

Severe Combined Immune Deficiency ◽

Dsb Repair ◽

G2 Checkpoint ◽

G2 Phase

Mice homozygous for the scid (severe combined immune deficiency) mutation are defective in the repair of DNA double-strand breaks (DSBs) and are consequently very X-ray sensitive and defective in the lymphoid V(D)J recombination process. Recently, a strong candidate for the scid gene has been identified as the catalytic subunit of the DNA-dependent protein kinase (DNA-PK) complex. Here, we show that the activity of the DNA-PK complex is regulated in a cell cycle-dependent manner, with peaks of activity found at the G1/early S phase and again at the G2 phase in wild-type cells. Interestingly, only the deficit of the G1/early S phase DNA-PK activity correlated with an increased hypersensitivity to X-irradiation and a DNA DSB repair deficit in synchronized scid pre-B cells. Finally, we demonstrate that the DNA-PK activity found at the G2 phase may be required for exit from a DNA damage-induced G2 checkpoint arrest. These observations suggest the presence of two pathways (DNA-PK-dependent and -independent) of illegitimate mammalian DNA DSB repair and two distinct roles (DNA DSB repair and G2 checkpoint traversal) for DNA-PK in the cellular response to ionizing radiation.

Download Full-text

Artemis-dependent DNA double-strand break formation at stalled replication forks

Cancer Science ◽

10.1111/cas.12144 ◽

2013 ◽

Vol 104 (6) ◽

pp. 703-710 ◽

Cited By ~ 12

Author(s):

Junya Unno ◽

Masatoshi Takagi ◽

Jinhua Piao ◽

Masataka Sugimoto ◽

Fumiko Honda ◽

...

Keyword(s):

Strand Break ◽

Double Strand Break ◽

Replication Forks ◽

Dna Double Strand Break ◽

Stalled Replication Forks ◽

Break Formation

Download Full-text

Mitotic replisome disassembly in vertebrates

10.1101/418368 ◽

2018 ◽

Author(s):

Sara Priego Moreno ◽

Rebecca M. Jones ◽

Divyasree Poovathumkadavil ◽

Agnieszka Gambus

Keyword(s):

Cell Cycle ◽

Dna Replication ◽

Ubiquitin Ligase ◽

S Phase ◽

Replication Forks ◽

Replication Machinery ◽

Replicative Helicase ◽

Egg Extract ◽

Eukaryotic Dna ◽

Higher Eukaryotes

ABSTRACTRecent years have brought a breakthrough in our understanding of the process of eukaryotic DNA replication termination. We have shown that the process of replication machinery (replisome) disassembly at the termination of DNA replication forks in S-phase of the cell cycle is driven through polyubiquitylation of one of the replicative helicase subunits Mcm7. Our previous work in C.elegans embryos suggested also an existence of a back-up pathway of replisome disassembly in mitosis. Here we show, that in Xenopus laevis egg extract, any replisome retained on chromatin after S-phase is indeed removed from chromatin in mitosis. This mitotic disassembly pathway depends on formation of K6 and K63 ubiquitin chains on Mcm7 by TRAIP ubiquitin ligase and activity of p97/VCP protein segregase. The mitotic replisome pathway is therefore conserved through evolution in higher eukaryotes. However, unlike in lower eukaryotes it does not require SUMO modifications. This process can also remove any helicases from chromatin, including “active” stalled ones, indicating a much wider application of this pathway than just a “back-up” for terminated helicases.

Download Full-text

Double-Strand Break Generation under Deoxyribonucleotide Starvation in Escherichia coli

Journal of Bacteriology ◽

10.1128/jb.00411-07 ◽

2007 ◽

Vol 189 (15) ◽

pp. 5782-5786 ◽

Cited By ~ 30

Author(s):

Estrella Guarino ◽

Israel Salguero ◽

Alfonso Jiménez-Sánchez ◽

Elena C. Guzmán

Keyword(s):

Escherichia Coli ◽

Replication Fork ◽

Thermal Inactivation ◽

Strand Break ◽

Double Strand Break ◽

Replication Forks ◽

Thymine Starvation ◽

Hydroxyurea Treatment ◽

Deoxynucleoside Triphosphate ◽

Stalled Replication Forks

ABSTRACT Stalled replication forks produced by three different ways of depleting deoxynucleoside triphosphate showed different capacities to undergo “replication fork reversal.” This reaction occurred at the stalled forks generated by hydroxyurea treatment, was impaired under thermal inactivation of ribonucleoside reductase, and did not take place under thymine starvation.

Download Full-text

Evidence for Biased Holliday Junction Cleavage and Mismatch Repair Directed by Junction Cuts during Double-Strand-Break Repair in Mammalian Cells

Molecular and Cellular Biology ◽

10.1128/mcb.21.10.3425-3435.2001 ◽

2001 ◽

Vol 21 (10) ◽

pp. 3425-3435 ◽

Cited By ~ 13

Author(s):

Mark D. Baker ◽

Erin C. Birmingham

Keyword(s):

Homologous Recombination ◽

Mismatch Repair ◽

Mammalian Cells ◽

Strand Break ◽

Double Strand Break ◽

Double Strand Break Repair ◽

Holliday Junction ◽

Chromosomal Dna ◽

Dna Breaks ◽

Break Repair

ABSTRACT In mammalian cells, several features of the way homologous recombination occurs between transferred and chromosomal DNA are consistent with the double-strand-break repair (DSBR) model of recombination. In this study, we examined the segregation patterns of small palindrome markers, which frequently escape mismatch repair when encompassed within heteroduplex DNA formed in vivo during mammalian homologous recombination, to test predictions of the DSBR model, in particular as they relate to the mechanism of crossover resolution. According to the canonical DSBR model, crossover between the vector and chromosome results from cleavage of the joint molecule in two alternate sense modes. The two crossover modes lead to different predicted marker configurations in the recombinants, and assuming no bias in the mode of Holliday junction cleavage, the two types of recombinants are expected in equal frequency. However, we propose a revision to the canonical model, as our results suggest that the mode of crossover resolution is biased in favor of cutting the DNA strands upon which DNA synthesis is occurring during formation of the joint molecule. The bias in junction resolution permitted us to examine the potential consequences of mismatch repair acting on the DNA breaks generated by junction cutting. The combination of biased junction resolution with both early and late rounds of mismatch repair can explain the marker patterns in the recombinants.

Download Full-text