Daniel Kaplan is an Associate Professor in the Department of Biomedical Sciences at Florida State University College of Medicine. Dr. Kaplan directs a lab that studies DNA replication and genome maintenance. Dr. Kaplan is the Block Director for Foundations 2: Molecules to Medicine, and Dr. Kaplan also teaches microbiology to medical student throughout the first two years of the curriculum. In addition, Dr. Kaplan teaches an outreach program on DNA replication to Leon County high school students and teachers, as part of a program funded by the National Science Foundation.
Daniel Kaplan joined the Florida State University College of Medicine in 2012 as an Associate Professor of Biomedical Sciences. Prior to joining FSU, Dr. Kaplan was an Assistant Professor at Vanderbilt University, where he led a team that published primary research articles that investigated how DNA replication is initiated. As a post-doctoral fellow, Daniel Kaplan worked with National Academy of Science member Mike O’Donnell at Rockefeller University and published manuscripts on the function and mechanism of replication fork helicases. Two of these manuscripts (Mol Cell 2002 and Mol Cell 2004) have been cited by the Faculty of 1000 website. In graduate school, Daniel Kaplan worked with Nobel Laureate Thomas Steitz at Yale University to determine how the bacterial replication fork helicase, DnaB, modulates DNA structure. Dr. Kaplan attended Yale Medical School and received training in cancer research before entering graduate school.
Ph.D. Yale University, New Haven, Connecticut
September 1994 to August 2000
Degree Date: December 15, 2000
Molecular Biophysics and Biochemistry
Dissertation Title: Mechanistic Studies of DnaB Helicase
Mentor’s Name: Thomas A. Steitz
Yale University School of Medicine, New Haven, Connecticut
September 1988 to January 1992
Passed Part I of National Medical Board Examination in 1990.
Completed 2 ½ years of medical school training and a one-year research NIH Cancer Research Fellowship.
Bachelor of Arts, University of Virginia, Charlottesville, Virginia
September 1984 to May 1988
Degree Date: May 1988
(2012- present) Member of 1st and 2nd year course directors committee
(2012- present) Member of Florida State University grant review workshop (2013- present) Co-founder of Florida State University Nucleic Acids Research Group (2014- present) Member of faculty council
Awarded tenure at the Florida State University College of Medicine (August 2015).
Invited to write a review for a special molecular motors issue of Frontiers in Biosciences, in the Encyclopedia of Bioscience (2012 publication).
Invited to submit an article to the journal Methods (2010 publication).
Invited to submit an article to the journal Methods in Molecular Biology (2010 publication).
Awarded the Scaringe Prize (2004 Gordon Research Conference: “Nucleic Acids”, Newport, Rhode Island, June 2004).
Leon and Toby Cooperman Fellow of the Damon Runyon Cancer Research Foundation (2001 to 2004).
Distinguished Doctoral Thesis (Awarded by Yale University Degree Committee, December 2000).
Howard Hughes Medical Institute Predoctoral Fellow (1995 to 2000).
Honor Student at an International Achievement Summit (hosted by the American Academy of Achievement, London, England, October 2000).
Nominated for Harold M. Weintraub Graduate Student Award (2000).
Poster Presentation Award (Yale University Graduate Student Research Symposium, February 1999).
Teaching Prize (Yale University Department of Molecular Biophysics and Biochemistry, June 1998).
Grade of Honors in 8 out of 9 Courses (Yale University Graduate School, 1994-1995, other grade is high pass).
NIH Cancer Research Fellow (Yale University School of Medicine, 1990-1991).
Student Editor of Yale Journal of Biology and Medicine (1989-1991).
Graduates with honors from University of Virginia.
Phi Beta Kappa
Intermediate Honors (awarded to second year students with high academic achievement).
Echols Scholar (awarded to incoming students of exceptional promise).
American Society for Biochemistry and Molecular Biology
American Association for the Advancement of Science
DNA replication and Genome Maintenance
DNA replication is a key process in living cells that is important for cell growth and division. Disruption of DNA replication in higher organisms may lead to genome instability, which may ultimately result in a disease state such as cancer. Our lab is principally interested in how cells replicate their genomic DNA in eukaryotic cells in order to maintain faithful inheritance of the genome.
DNA replication is initiated by chromosomal unwinding, a process wherein the replication fork helicase separates the duplex genomic DNA into single strands. The single stranded DNA can then serve as a template for DNA polymerases to carry out faithful replication of the genome. The replication fork helicase in eukaryotes is composed of the Mcm2-7 heterohexamer, Cdc45, and the tetrameric GINS complex. The Mcm proteins serve as useful makers for cancer, and understanding how these proteins function may lead to further advancement in the development of cancer prognostic and diagnostic tools. Understanding how the replication fork helicase initiates unwinding in eukaryotic cells to initiate DNA replication is a focus of study in our laboratory. We are also interested in determining how unwinding at a replication fork is coupled with DNA duplication, and the cellular polymerases pol alpha, pol delta, and pol epsilon catalyze synthesis of DNA.
The initiation of DNA replication in eukaryotes is regulated by cellular kinases, including the Dbf4-Cdc7 kinase, and the S-phase cyclin-dependent kinase. These kinases act upon components of the replication fork to activate the replication fork helicase. These kinases are currently the targets for chemotherapeutic agents because of their central role in controlling the initiation of DNA replication. Four additional proteins are required for DNA replication initiation in budding yeast cells, Sld2, Sld3, Dpb11, and Mcm10. We are investigating the mechanism of action of these four proteins because they are central to understanding how DNA replication is initiated.
Our investigations will yield essential information about how DNA replication is initiated and regulated in eukaryotic cells. Importantly, we will provide a detailed, mechanistic, and dynamic understanding of how the protein machines that initiate replication function in a cell. The advances that we make will likely lead to improvements in the treatment of cancer. Some of the proteins that we study are also involved in the DNA damage response. Thus, an additional emphasis is to determine how the DNA damage response functions to allow cells to survive upon challenge by agents that are toxic to DNA.
Articles (in print or accepted) in refereed journals:
C Eielson, D Kaplan, MA Mitnick, I Paliwal, K Insogna. “Estrogen modulates parathyroid hormone-induced fibronectin production in human and rat osteoblast-like cells.” Endocrinology 135: 1639-1644 (1994). PMID: 8647916.
DL Kaplan, WF Boron. “Long-term expression of c-H-ras stimulates Na-H and Na+-dependent Cl-HCO3 exchange in NIH 3T3 fibroblasts.” J. Biol. Chem. 269: 4116-4124 (1994). PMID: 8307971.
DL Kaplan, CM Eielson, MC Horowitz, KL Insogna, EC Weir. “Tumor necrosis factor-alpha induces transcription of the colony-stimulating factor 1 gene in murine osteoblasts.” J. Cell Physiol. 168: 199-208 (1996). PMID: 8647916.
DL Kaplan, TA Steitz. “DnaB from Thermus aquaticus Unwinds Forked Duplex DNA with an Asymmetric Tail Length Dependence.” J. Biol. Chem. 274: 6889-6897 (1999). PMID: 10066742
DL Kaplan. “The 3’-tail of a Forked-Duplex Sterically Determines Whether One or Two DNA Strands Pass Through the Central Channel of a Replication Fork Helicase.” J. Mol. Biol. 301: 285-299 (2000). PMID: 10926510.
DL Kaplan, M O'Donnell. “DnaB Drives DNA Branch Migration and Dislodges Proteins While Encircling Two DNA Strands.” Mol. Cell 10: 647-657 (2002). PMID: 12408831.
DL Kaplan and M O’Donnell. “Rho Factor: Transcription Termination in Four Steps.” Curr. Biol. 13: R714-R716 (2003). PMID: 13678611.
DL Kaplan, MJ Davey, M O'Donnell. “Mcm4,6,7 Uses ‘Pump in Ring’ Mechanism to Unwind DNA by Steric Exclusion and Actively Translocate Along a Duplex.” J. Biol. Chem. 278: 49171-49182 (2003). PMID: 13679365.
DL Kaplan and M O'Donnell. “Twin DNA Pumps of a Hexameric Helicase Provide Power to Simultaneously Melt Two Duplexes.” Mol. Cell 15: 453-465 (2004). PMID: 15304224.
DL Kaplan and M O'Donnell. “RuvA Is a Sliding Collar that Protects Holliday Junctions from Unwinding while Promoting Branch Migration.” J. Mol. Biol. 355: 473-490 (2006). PMID: 16324713.
DL Kaplan. “Replication Termination: Mechanism of Polar Arrest Revealed.” Curr. Biol. 16: R684-686 (2006). PMID: 16950091.
DM Kanter, I Bruck, and DL Kaplan. “Mcm subunits can assemble into two different active unwinding complexes.” J. Biol. Chem. 283:31172-1182 (2008). PMID: 18801730.
DL Kaplan and D Bastia. “Mechanisms of polar arrest of a replication fork.” Mol. Microbiol. 72:27-285 (2009). PMID: 19298368.
I Bruck and D Kaplan. “Dbf4-Cdc7 phosphorylation of Mcm2 is required for cell growth.” J. Biol. Chem. 284:28823-31 (2009). PMID: 19692334.
DL Kaplan and I Bruck. “Methods to study kinase regulation of the replication fork helicase.” Methods. 51:358-62 (2010). PMID: 20170732.
CE Wickersham, KJ Cash, SH Pfeil, I Bruck, DL Kaplan, KW Plaxco, and EA Lipman. “Tracking a molecular motor with a nanoscale optical encoder.” Nano Lett. 10:1022-1027 (2010). PMID: 20121107.
DL Kaplan and I Bruck. “Methods to study how replication fork helicases unwind DNA.” Methods Mol Biol. 587:127-35 (2010). PMID: 20225146
N Ribeck, DL Kaplan, I Bruck, OA Saleh, “DnaB helicase activity is modulated by DNA geometry and force.” Biophys J. 99:2170-9 (2010). PMID: 20923651
DM Kanter and DL Kaplan, "Sld2 binds to origin ssDNA and stimulates DNA annealing." Nucleic Acids Res. 39: 2580-2592 (2011) PMID: 21109535.
I. Bruck and DL Kaplan, "GINS and Sld3 compete with one another for Mcm2-7 and Cdc45 binding." J. Biol. Chem. 286:14157-67 (2011) PMID: 21362622.
I. Bruck and DL Kaplan, "Origin Single-stranded DNA Releases Sld3 Protein from the Mcm2-7 Complex, Allowing the GINS Tetramer to Bind the Mcm2-7 Complex." J. Biol. Chem. 286:18602-13 (2011) PMID: 21460226.
I. Bruck, DM Kanter, and DL Kaplan "Enabling Association of the GINS Protein Tetramer with the Mini Chromosome Maintenance (Mcm)2-7 Protein Complex by Phosphorylated Sld2 Protein and Single-stranded Origin DNA." J. Biol. Chem. 286:36414-26 (2011) PMID: 21868389.
AN Suhasini, JA Sommers, S Yu, Y Wu, T Xu, Z Kelman, DL Kaplan, and RM Brosh, Jr. “DNA Repair and Replication Fork Helicases are Differentially Affected by an Alkyl Phosphotriester Lesion.” J. Biol. Chem. 287:19188-98 (2012) PMID: 22465574.
DL Kaplan, OA Saleh, and N Ribeck “Single-molecule and bulk approaches to the DnaB replication fork helicase.” Front. Biosci. 18: 224-40 (2013) PMID: 23276919.
I Bruck and DL Kaplan, “Cdc45-ssDNA interaction is important for stalling the helicase during replication stress.” J Biol Chem. 288: 7550-63 (2013) PMID: 23382391.
Bruck, I., and Kaplan, D. The replication initiation protein sld2 regulates helicase assembly. J Biol Chem 289, 1948-1959. (2014) PMID: 24307213.
Bharti SK, Sommers JA, Zhou J, Kaplan DL, Spelbrink JN, Mergny JL, Brosh RM Jr. DNA sequences proximal to human mitochondrial DNA deletion breakpoints prevalent in human disease form G-quadruplexes, a class of DNA structures inefficiently unwound by the mitochondrial replicative Twinkle helicase. The Journal of biological chemistry. 289(43):29975-93 (2014). PMID: 25193669.
Dhingra, N., Bruck, I., Smith, S., Ning, B., and Kaplan, D. Dpb11 helps control assembly of the Cdc45-Mcm2-7-GINS replication fork helicase. J Biol Chem 290, 7586-7601. (2015) PMID: 25659432.
Bruck, I., and Kaplan, D. The replication initiation protein Sld3/Treslin orchestrates the assembly of the replication fork helicase during S phase. J Biol Chem 290, 27414-27424. (2015) PMID: 26405041.
Bruck, I., and Kaplan, D. L. The Dbf4-Cdc7 kinase promotes Mcm2-7 ring opening to allow for single-stranded DNA extrusion and helicase assembly. J. Biol Chem. 290, 1210-1221. (2015) PMID: 25471369.
Bruck, I., and Kaplan, D. Conserved mechanism for coordinating replication fork helicase assembly with phosphorylation of the helicase. Proc Natl Acad Sci U S A. 112, 11223-11228. (2015) PMID: 26305950.
Bruck I, Perez-Arnaiz P, Colbert MK, Kaplan DL. Insights into the Initiation of Eukaryotic DNA Replication. Nucleus. (2015); 6(6):449-54. PMID: 26710261
Perez-Arnaiz, P., Bruck, I., and Kaplan, D. Mcm10 coordinates the timely assembly and activation of the replication fork helicase. Nucleic Acids Res. 44, 315-329. (2016) PMID: 26582917.
2. Book Chapters:
Helicases, Edited by Mohamed M. Abdelhaleem, Published by Humana Press, Inc., Series: Methods in Molecular Biology, Volume No. 587, ISBN: 9781603273541, Chapter title: “Methods to Study How Replication Fork Helicases Unwind DNA”, authored by Daniel L. Kaplan and Irina Bruck, pages 127-136 (2010).
N Dhingra, DL Kaplan. The Initiation of DNA Replication in Eukaryotes. pages 1-21 from the book: Introduction to Eukaryotic DNA Replication Initiation. edited by Daniel L. Kaplan (2016)