James P. Carney, Ph.D. Assistant Professor of Radiation Oncology
Department:
Radiation Oncology
UMGCC Research Program:
Molecular and Structural Biology Program
Education/Training:
College Degree:
B.S. Chemistry/Education, Niagara University
Medical Degree:
Ph.D. Biochemistry, Loyola University of Chicago
Post Doctoral
Degree:
University of California, Berkeley
University of California, San Francisco
Lawrence Berkeley National Laboratory
Contact
Information:
Mailing Address:
University of Maryland School of Medicine
The Radiation Oncology Research Laboratory
Bressler 6-011
655 West Baltimore St.
Baltimore, MD 21201
Email:
jcarney@som.umaryland.edu
Phone:
410-706-4276
Fax:
410-706-6138
Research Interests:
Our laboratory is primarily interested in the structure and function of multi-protein complexes involved in the cellular response to DNA damage. Our model of choice for this work is the Mre11/Rad50/Nbs1 complex (or Mre11 complex). This complex is responsible for repair of DNA double-stand breaks, detection of DNA damage and telomere maintenance as well as functions in meiosis. In all of these various functions a mechanistic understanding of how the Mre11 complex works has not been developed. Mre11 has been demonstrated to be a 3 to 5 nuclease and Rad50 is an ATP-stimulated DNA binding protein. The Nbs1 protein is somewhat more enigmatic as it does not appear to have any enzymatic activities although it does stimulate the activities of Mre11 and Rad50. Furthermore, genetic studies in humans have demonstrated that mutations of the gene coding for Nbs1 results in the disease Nijmegen breakage syndrome while partial loss of function mutations in Mre11 lead to the disorder ataxia telangiectasia-like disorder (ATLD). Both of the diseases share a number of similarities including chromosomal fragility, impaired immune function, radiation sensitivity and increased cancer incidence. Given the radiation sensitivity and chromosomal fragility associated with defects in the Mre11 complex it is clear that the complex has a critical role in the cellular response to DNA damage, in particular DNA double-strand breaks. In order to understand mechanism of the Mre11 complex function in all of its various in vivo roles we are investigating the structural and functional attributes that govern the biochemical activities. It is our hypothesis that there are a limited number of activities of the Mre11 complex that are utilized for all of its in vivo functions.
In order to investigate this hypothesis we are pursuing several lines of experimentation. Our primary approach is the expression, purification and characterization of the subunits of the human Mre11 complex using an insect cell/baculovirus system. Coupled with the biochemical analysis of the Mre11 complex we are pursuing structural studies in collaboration with the laboratory of John Tainer. The ultimate goal of this work is the integration of in vivo genetic studies, in vitro biochemical analysis and state of the art structural biology to come to an enhanced understanding of the function of the Mre11 complex as well as utilize this information for the design of therapeutics for cancer treatments.
Publications:
Arthur, L.M., Gustausson, K., Hopfner, K-P., Carson, C.T., Stracker, T.H., Karcher, A., Felton, D., Weitzman, M.D., Tainer, J.A. and Carney, J.P. (2004) Structural and Functional Analysis of Mre11-3. Nucleic Acids Research. 32: 1886-1893.
DeJager, M., Trujillo, K., Sung, P., Hopfner, K-P., Carney, J.P., Tainer, J.A., Connelly, J.C.,Leach D.R.F., Haering, C., Nasmyth, K., Kannaar, R. and Wyman, C. (2004) Differential arrangement of conserved building blocks among homologs of the Rad50/Mre11 DNA repair complex. Journal of Molecular Biology. 339: 937-949.
Cheng, W-H., von Kobbe, C., Opresko,P.L., Arthur, L.M., Komatsu, K., Seidman, M.M., Carney, J.P. and Bohr, V.A. (2004) Linkage between werner syndrome protein and the Mre11 complex via Nbs1. Journal of Biological Chemistry. 279: 21169-21176.
Arthur, L.M., Gustausson, K., Hopfner, K-P., Carson, C.T., Stracker, T.H., Karcher, A., Felton, D., Weitzman, M.D., Tainer, J.A. and Carney, J.P. (2004) Structural and Functional Analysis of Mre11-3. Nucleic Acids Research. 32: 1886-1893.
Manzan, A., Pfeiffer, G., Hefferin, M.L., Lang, C.E., Carney, J.P. and Hopfner, K-P. (2004) MlaA, a hexameric ATPase linked to the Mre11 complex in archaeal genomes. EMBO Reports 5:5459
Hopfner, K.P., Craig, L., Moncalian, G., Zinkel, R.A., Takehiko, U., Owen, B.L., Karcher, A., Henderson, B., Bodmer, J-L., McMurray, C.T., Carney, J.P., Petrini, J.H.J. and Tainer, J.A. (2002) Rad50 Zn hook: a novel structure joining Mre11 complexes in DNA recombination and repair. Nature 418:562-6.
Hopfner, K.P., Karcher, A., Craig, L., Woo,T.T., Carney, J.P. and Tainer, J.A. (2001) Structural biochemistry and interaction architecture of the DNA double-strand break repair Mre11 nuclease and Rad50 ATPase. Cell 105:473-485.
Hopfner, K.P., Karcher, A., Fairley, C., Tainer, J.A. and Carney, J.P. (2000) Mre11 and Rad50 from Pyrococcus furiosus: Cloning and biochemical characterization reveal an evolutionarily conserved multiprotein machine. J. Bacteriology 182:6036-6041.
Hopfner, K-P., Karcher, A., Shin, D.S., Craig, L., Arthur, L.M., Carney, J.P. and Tainer, J.A. (2000) Structural biochemistry of Rad50 ATPase: ATP-driven cooperativity and allosteric control in DNA double-strand break repair and the ABC ATPase superfamily. Cell 101: 789-800.
Carney, J.P. (1999) Chromosomal breakage syndromes. Current Opinion in Immunology. 11: 443-447.
Carney, J.P., Maser, R., Olivares, H., Davis, E.M., Le Beau, M., Yates, J.R.III, Hays, L., Morgan, W.F., and Petrini, J.H.J. (1998) The hMre11/hRad50 protein complex and Nijmegen breakage syndrome: Linkage of double-strand break repair to the cellular DNA damage response. Cell 93: 477-486.
