DNA replication in eukaryotic cells is tightly controlled so that the genome is replicated once and only once per cell cycle. Such a mechanism is extremely important for faithful transmission of genetic information from one generation to the next. Over-replication of chromosomal DNA will result in genetic instability, which is often associated with human diseases, such as cancer. Our long-term objectives are to understand how DNA rereplication is prevented in mammalian cells and how loss of DNA replication control would lead to genome instability and tumorigenesis. One key to the control of DNA replication initiation is the tightly regulated assembly of pre- replication complexes (pre-RCs) at replication origins by the licensing control mechanism. We showed that when the licensing control is compromised, the ATR-mediated checkpoint is activated and plays a critical role in the suppression of DNA rereplication. Our recent studies further demonstrate that DNA repair machineries are actively involved in removing over-replicated DNA regions and repairing rereplication-associated DNA lesions. We propose that both S-phase checkpoint and DNA repair functions are critical for the maintenance of genome stability when the licensing control is impaired. In this proposal, we will investigate the mechanisms underlying the S-phase checkpoint control and DNA repair activities in the suppression of DNA rereplication and in the repair of rereplication-associated DNA lesions. First, we will study the biological importance of the communication between the S-phase checkpoint and the replication licensing control through a direct interaction of checkpoint proteins with the licensing factor Cdt1. Second, we will investigate the role of DNA repair mechanisms in the removal of rereplicated DNA and in the repair of DNA double-strand breaks (DSBs) that arise during DNA rereplication. We will also define the repair pathways which are used to repair DSBs during DNA rereplication. Third, we will probe the role of the Mre11/Rad50/Nbs1 complex in the suppression of DNA rereplication and investigate the mechanisms underlying this function. DNA rereplication would inevitably lead to genome instability, which is an integral aspect of the malignant phenotype. Recent observations that rereplication or unscheduled DNA replication is induced at the initial stages of tumorigenesis highlight the importance of replication control in the prevention of cancer. Understanding how rereplication is prevented in mammalian cells will shed light on the cellular mechanisms which govern genome stability and improve our understanding of cancer etiology.

Public Health Relevance

DNA rereplication or over-replication of chromosomal DNA often contributes to genome instability, which is highly associated with cancer development. Therefore, understanding how DNA rereplication is suppressed in mammalian cells is of great importance for clarifying the mechanisms underlying the prevention of tumorigenesis. The proposed studies will shed light on cancer etiology and will ultimately help develop new therapeutic interventions for human diseases associated with genome instability and cancer.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM080677-04
Application #
8535163
Study Section
Cancer Etiology Study Section (CE)
Program Officer
Reddy, Michael K
Project Start
2010-09-15
Project End
2014-08-31
Budget Start
2013-09-01
Budget End
2014-08-31
Support Year
4
Fiscal Year
2013
Total Cost
$344,700
Indirect Cost
$163,183
Name
Scripps Research Institute
Department
Type
DUNS #
781613492
City
La Jolla
State
CA
Country
United States
Zip Code
92037
Wei, Na; Shi, Yi; Truong, Lan N et al. (2014) Oxidative stress diverts tRNA synthetase to nucleus for protection against DNA damage. Mol Cell 56:323-32
Wang, Hailong; Li, Yongjiang; Truong, Lan N et al. (2014) CtIP maintains stability at common fragile sites and inverted repeats by end resection-independent endonuclease activity. Mol Cell 54:1012-21
Truong, Lan N; Li, Yongjiang; Shi, Linda Z et al. (2013) Microhomology-mediated End Joining and Homologous Recombination share the initial end resection step to repair DNA double-strand breaks in mammalian cells. Proc Natl Acad Sci U S A 110:7720-5
Truong, Lan N; Wu, Xiaohua (2011) Prevention of DNA re-replication in eukaryotic cells. J Mol Cell Biol 3:13-22