A hallmark of cancer and other diseases are high rates of mutation and genome instability, which are believed to be the driving force for disease causing genetic changes. The DMA damage checkpoint is important for the suppression of mutations and genome instabilities, and the regulation of many cellular responses to DMA damage. While an increasing number of genes have been identified to function in the DMA damage checkpoint in human and mutations in some checkpoint genes are linked to cancers, fundamental questions of how the DNA damage checkpoint is regulated remain far from being understood. An important reason for using model experimental systems is to rapidly obtain experimental insights into biological problems that can be applied to problems of human diseases, especially for highly conserved pathways like the DNA damage checkpoint. Yeast, in particular, offers unparalleled advantages in yeast genetics and short generation time. Basic studies in yeast have contributed significantly to the understanding of the DNA damage checkpoint. In the proposed basic studies, we are primarily interested in two basic mechanistic questions about the DNA damage checkpoint. First, how is the DNA damage checkpoint activated? Second, how are various DNA damage checkpoint pathways linked to various DNA damage responses? Accordingly, there are three complementary aims in this proposal. First, we will examine the genetic pathways for Rad53 and Dun1 activation using SmM as a substrate. Second, we will dissect the role of Dun1 phosphorylation in its activation and characterize the roles of Dun1 phosphorylation in the functions of Dun1, including the regulation of dNTP levels, telomere maintenance and suppression of gross chromosome rearrangements. Third, we will use a newly developed Rad53- Dun1-Sml1 system to dissect how Mec1, Tell and Rad53 are activated via biochemical reconstitution and genetic analysis. We will use purified proteins to reconstitute the activation of the Rad53-Dun1-Sml1 system, and further use this system to study the regulation of Mec1 and Tell. Our long-term goal is to understand the regulation of the DNA damage checkpoint kinases and their functions in the regulation of cell growth and suppression of genome instability.

National Institute of Health (NIH)
National Institute of General Medical Sciences (NIGMS)
Research Project (R01)
Project #
Application #
Study Section
Molecular Genetics A Study Section (MGA)
Program Officer
Hagan, Ann A
Project Start
Project End
Budget Start
Budget End
Support Year
Fiscal Year
Total Cost
Indirect Cost
Ludwig Institute for Cancer Research Ltd
La Jolla
United States
Zip Code
Liang, Jason; Suhandynata, Raymond T; Zhou, Huilin (2015) Phosphorylation of Sae2 Mediates Forkhead-associated (FHA) Domain-specific Interaction and Regulates Its DNA Repair Function. J Biol Chem 290:10751-63
Albuquerque, Claudio P; Wang, Guoliang; Lee, Nancy S et al. (2013) Distinct SUMO ligases cooperate with Esc2 and Slx5 to suppress duplication-mediated genome rearrangements. PLoS Genet 9:e1003670
Wang, Guoliang; Tong, Xiangyan; Weng, Stephanie et al. (2012) Multiple phosphorylation of Rad9 by CDK is required for DNA damage checkpoint activation. Cell Cycle 11:3792-800
Ling, Shuo-Chien; Albuquerque, Claudio P; Han, Joo Seok et al. (2010) ALS-associated mutations in TDP-43 increase its stability and promote TDP-43 complexes with FUS/TLS. Proc Natl Acad Sci U S A 107:13318-23
Chen, Sheng-hong; Albuquerque, Claudio P; Liang, Jason et al. (2010) A proteome-wide analysis of kinase-substrate network in the DNA damage response. J Biol Chem 285:12803-12
Zhou, Huilin; Albuquerque, Claudio P; Liang, Jason et al. (2010) Quantitative phosphoproteomics: New technologies and applications in the DNA damage response. Cell Cycle 9:3479-84
Dippold, Holly C; Ng, Michelle M; Farber-Katz, Suzette E et al. (2009) GOLPH3 bridges phosphatidylinositol-4- phosphate and actomyosin to stretch and shape the Golgi to promote budding. Cell 139:337-51
Tang, Tingdong; Zheng, Bin; Chen, Sheng-Hong et al. (2009) hNOA1 interacts with complex I and DAP3 and regulates mitochondrial respiration and apoptosis. J Biol Chem 284:5414-24
Saito, Kota; Chen, Mei; Bard, Fred et al. (2009) TANGO1 facilitates cargo loading at endoplasmic reticulum exit sites. Cell 136:891-902
Wagner, Michelle V; Smolka, Marcus B; de Bruin, Rob A M et al. (2009) Whi5 regulation by site specific CDK-phosphorylation in Saccharomyces cerevisiae. PLoS One 4:e4300

Showing the most recent 10 out of 17 publications