Anja K. Bielinsky, PhD
Professor, and Biophysics Department of Biochemistry, Molecular Biology
Professor, and Biophysics Department of Biochemistry, Molecular Biology
Associate Dean for Foundational Science, Medical School
Associate Director for Basic Shared Resources, Masonic Cancer Center (MCC)
Faculty, MS and PhD Programs in Pharmacology
Special Fellow, Leukemia and Lymphoma Society (1999-2001)
PhD, Heinrich Heine University, Düsseldorf, Germany, 1995
American Cancer Society Scholar (2002-2006),
Leukemia and Lymphoma Society Scholar (2008-2013)
Regulation of DNA replication, Cell cycle, Cancer
DNA replication is an integral part of the eukaryotic cell cycle, which coordinates the duplication of the entire genome during S phase. At the onset of S phase, origins of DNA replication are activated to "fire". Once DNA synthesis has been initiated, the cell is committed to undergo a complete round of division. Dysregulation of origin activation results, therefore, in uncontrolled cell proliferation, which is at the heart of diseases such as cancer. To initiate DNA replication, cells must assemble replication initiation complexes. Key components of these complexes are the minichromosome maintenance (Mcm) proteins. Mcm2-7 comprise the core of the replicative helicase and Mcm10 is a scaffold protein that links polymerization to DNA unwinding (it connects Mcm2-7 with Pol? in Figure 1).
Disruption of this coordination leads to replication fork stalling, commonly known as replication stress. The concept of replication stress-induced chromosome breakage is becoming increasingly important in cancer biology and the main focus of our laboratory is to understand the genome integrity network in eukaryotic cells. We are particularly interested in the role that Mcm10 plays to prevent double strand break formation during DNA replication. Multiple cancer-associated mutations have been mapped in Mcm10 (shown for the conserved core domain in Figure 2).
Another focus in our lab is the investigation of the S phase checkpoint and DNA damage tolerance pathways. All eukaryotic cells have evolved these pathways to respond to replication stress when replication forks encounter obstacles such as damaged DNA or other impediments that force them to slow down. The S phase checkpoint maintains the stability of replication forks under such circumstances, prevents replication fork collapse and thus chromosome breakage. S phase checkpoint genes act as an early cancer barrier in normal cells and loss-of-function mutations directly contribute to cancer development. The DNA damage tolerance pathways are activated in parallel to the S phase checkpoint and execute their function through ubiquitination of the replication clamp, proliferating cell nuclear antigen (PCNA). Ubiquitination acts as a flag to mark replication forks that are in trouble and induces the formation of complexes that allow replication forks to resume DNA synthesis.
- Alver, R.C., T. Zhang, A. Rajan, S. Das-Bradoo and A. K. Bielinsky, 2014. The N-terminus of Mcm10 is important for interaction with the 9-1-1 clamp and in resistance to DNA damage. Nucleic Acids Res. 42: 8389-8404. (pubmed)
- Becker, J.R., H.D. Nguyen, X. Wang and A. K. Bielinsky, 2014. Mcm10 deficiencycauses defective-replisome-induced mutagenesis and a dependency on postreplicative repair. Cell Cycle 13: 1737-1748. (pubmed)
- Thu, Y. M. and A. K. Bielinsky, 2014. Mcm10: one tool for all – integrity, maintenance and damage control. Semin. Cell. Dev. Biol. 30: 121-130. (pubmed)
- Ragland, R. L., S. Patel, R. Revard, K. Smith, A. A. Peters, A. K. Bielinsky, and E. J. Brown, 2013. RNF4 and PLK1 are required for replication fork collapse in ATR-deficient cells. Genes & Dev. 27: 2259-2273. (pubmed)
- Thu, Y. M. and A. K. Bielinsky, 2013. Enigmatic roles of Mcm10 in DNA replication, TIBS 38: 184-194. (pubmed).
- Nguyen, H. D., J. Becker, Y. M. Thu, M. Costanzo, E. N. Koch, S. Smith, K. Myung, C. L. Myers, C. Boone and A. K. Bielinsky, 2013. Unligated Okazaki fragments induce PCNA ubiquitination and a requirement for Rad59-dependent replication fork progression, PLoS ONE 8(6): e66379. (pubmed)
- Du, W., A. Josephrajan, S. Adhikary, T. Bowles, A. K. Bielinsky, and B. Eichman, 2013. Mcm10 self-association is mediated by an N-terminal coiled-coil domain, PLoS ONE 8(7):e70518. (pubmed)
- Haworth, J. C., Alver, R., Anderson, M. and Bielinsky, A. K. 2010. Ubc4 and Not4 regulate steady-state levels of DNA polymerase-alpha to promote efficient and accurate DNA replication, Molecular Biology of the Cell 21: 3205-3219. (pubmed)
- Nguyen, H. D. and Bielinsky, A. K. 2010. HDM2 ERKs PCNA, Journal of Cell Biology 190: 487-489. (pubmed)
- Alver, R. and Bielinsky, A. K. 2010. Termination at sTop2, Molecular Cell39: 487-489. (pubmed)
- Das-Bradoo, S., Nguyen, H. D. and Bielinsky, A. K. 2010. Damage-specific modification of PCNA, Cell Cycle 9: 3674-3679. (pubmed)
- Das-Bradoo, S., Nguyen, H. D., Ricke, R. M., Haworth, J. C. and Bielinsky, A. K. 2010. DNA ligase I deficiency triggers PCNA ubiquitination, Nature Cell Biology 12: 74-79. (pubmed)
- Das-Bradoo, S. and Bielinsky, A. K., 2009. Mapping replication initiation sites in eukaryotic genomes. Molecular Methods in DNA Replication (ed., John Walker and Jacob Dalgaard), Humana Press Inc.
- Raveendranathan, M. and Bielinsky, A. K., 2009. Analyzing origin activation patterns by copy-number change experiments. Molecular Methods in DNA Replication (ed., John Walker and Jacob Dalgaard),Humana Press Inc.
- Warren, E., Vaithiyalingam, S., Haworth, J., Greer, B., Smith, J.,Bielinsky, A. K., Chazin, W. and Eichman, B., 2008. Structural basis for DNA binding by replication initiator Mcm10, Structure (cover) 16:1892-1901. (pubmed)
- Bielinsky, A. K., 2007. Scarce but scary. Nature Genetics 39: 707-708. (pubmed)
- Chattopadhyay, S. and Bielinsky, A. K., 2007. Human Mcm10 regulates the catalytic subunit of DNA polymerase-alpha and prevents DNA damage during replication. Molecular Biology of the Cell 18: 4085-4095. (pubmed)
- Bolon, Y. T. and Bielinsky, A. K. 2006. The spatial arrangement of ORC binding modules determines the functionality of replication origins in budding yeast. Nucleic Acids Research 34:5069-5080. (pubmed)
- Raveendranathan, M., Chattopadhyay, S., Bolon, Y.-T., Haworth, J. C., Clarke, D. J. and Bielinsky, A. K. 2006. Genome-wide replication profiles of S phase checkpoint mutants reveal fragile sites in yeast,EMBO J. 25:3627-3639. (pubmed)
- Ricke, R. M. and Bielinsky, A. K. 2006. A conserved Hsp10-like domain in Mcm10 is required for the stabilization of DNA polymerase-alpha in budding yeast, Journal of Biological Chemistry 281:18414-18425. (pubmed)
- Das-Bradoo, S., Ricke, R. M. and Bielinsky, A. K. 2006. Interaction between PCNA and di-ubiquitinated Mcm10 is essential for cell growth in budding yeast, Molecular & Cell Biology 26:4806-4817. (pubmed)
- Bielinsky, A. K. and Raveendranathan, M. 2006. Encircled: large-scale purification of replication origins from mammalian chromosomes,Molecular Cell 21: 735-737. (Pubmed)
- Ricke, R. M. and Bielinsky, A. K. 2005. Easy detection of chromatin binding proteins by the histone association assay. Biological Procedures Online, 7:60-69. (Online citation)
- Ricke, R. M. and Bielinsky, A. K. 2004. Mcm10 regulates the stability and chromatin association of DNA polymerase-alpha. Molecular Cell, 16:173-185. (Medline citation)
- Bielinsky, A. K. 2003. Replication origins-why do we need so many?Cell Cycle 2:307-309. (Medline citation)
- Gerbi, S. A. and Bielinsky, A. K. 2002. DNA replication and chromatin.Curr. Opin. Genet. Dev. 12:243-248. Medline citation.
- Bielinsky, A. K., Blitzblau, H., Beall, E. L., Ezrokhi, M., Smith, H. S., Botchan, M. R., and Gerbi, S. A. 2001. Origin recognition complex binding to a metazoan replication origin. Current Biology 11:1427-1431.Medline citation.
- Bielinsky, A. K., and S. A. Gerbi, 2001. Where it all starts: eukaryotic origins of DNA replication. J. Cell Science 114: 643-651. Medline citation.
- Bielinsky, A. K., and S. A. Gerbi. 1999. Chromosomal ARS1 has a single leading strand start site. Molecular Cell 3:477-486. Medline citation.
- Gerbi, S. A., Bielinsky, A. K., Liang, C., Lunyak, V. V. and Urnov, F. D. 1999. Methods to map origins of replication in eukaryotes in "Eukaryotic DNA Replication: a practical approach" (ed., S. Cotterill),Oxford University Press, pp 1-41.
- Bielinsky, A. K., and S. A. Gerbi. 1998. Discrete start sites for DNA synthesis in the yeast ARS1 origin. Science 279:95-98. Medline citation.
- Gerbi, S. A., and Bielinsky, A. K. 1997. Replication initiation point mapping. Methods 13:271-280. Medline citation.