John J. Correia, Ph.D.

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The long range goal of our research is a detailed molecular understanding of microtubule assembly and disassembly. Microtubules are dynamic structures, undergoing spatial and temporal assembly and disassembly within the cell nucleus and cytoplasm. This dynamic behavior is under cell cycle regulation and is essential for normal cell function and growth. Microtubules are composed of tubulin and microtubule-associated proteins (MAPs). We are pursuing fundamental investigations of guanine nucleotide binding to tubulin, the effect of divalent cations on assembly and nucleotide hydrolysis, the role of MAPs in assembly and microtubule bundling, the importance of tubulin isotypes and their posttranslational modification in modulating microtubule dynamics and the interaction of anticancer drugs with microtubules and tubulin isotypes. Our current focus is on the vinca alkaloid-induced self-association of tubulin. We are comparing different vincas to evaluate the relationship between chemical modifications, tubulin spiral assembly and antimitotic efficacy. A knowledge of the energetics of these processes is essential to an understanding of microtubule dynamics and drug cytotoxicity [See the Current Projects page.]. Methods of analysis include: analytical ultracentrifugation, electron microscopy, DIC video- enhanced microscopy, light scattering, affinity purification, western blotting and computer modeling. The conclusions from these studies are relevant to the thermodynamics of microtubule assembly, the mechanisms of axonal transport, mitosis and morphogenesis, as well as potentially improving the therapeutic efficacy of antimitotic drugs in controlling and curing cancer. These studies are being extended to other drugs like Dolastatin 10 which also induce tubulin self-association and have therapeutic potential.

In addition, a number of collaborative projects are investigating the hydrodynamics and self-association of macromolecules by analytical ultracentrifugation. The systems being looked at include kinesin and NCD motor domains, B23 (a nucleolar protein), histone (H1)-DNA interactions, and SMAD proteins [See the Current Projects page.]. We are currently focusing on the development of analysis tools for investigating mixed-association, particularly the formation of 1:1 and 2:1 complexes of tubulin heterodimers with motor domain monomers and dimers. These experiments are important because they independently verify the affinity of motor domains, particularly single head or monomeric constructs, for tubulin, and allow for the dissection of cooperative interactions and the dependence upon both GXP and AXP nucleotides in the absence of processive ATP hydrolysis. [link to Kinesin home page]

This work also serves as the foundation for quantitative studies of tubulin interactions with other proteins like Kin-I, Clip-170 and OP18 that are involved in the regulation of microtubule dynamics and cell cycle. These studies are probing the role of anti-mitotic drugs in disrupting this regulation and will potentially lead to the development of new drugs for new sites of action.

The analytical ultracentrifugation is conducted in the UMMC Analytical Ultracentrifuge Facility.

Recent Publications

[or Link to Correia JJ PubMed Search]

Correia, J.J. and Chaires, J.B. Analysis of drug-DNA binding isotherms: a Monte Carlo approach. Methods Enzymol. 240: 593-614, 1994.

Lobert, S., Isern, N., Hennington, B.S. and Correia, J.J. The interaction of tubulin and microtubule proteins with vanadate oligomers. Biochemistry 33:6244-6252, 1994.

Correia, J.J., Gilbert, S.P., Moyer, M.L. and Johnson, K.A. Sedimentation studies on the Kinesin head domain constructs K401, K366, and K341. Biochemistry 34: 4898-4907, 1995.

Lobert, S., Frankfurter, A. and Correia, J.J. Binding of vinblastine to phosphocellulose-purified and αβ-Class III tubulin: the role of nucleotides and β-tubulin isotypes. Biochemistry. 34:8050-8060, 1995.

Rosenfeld, S.S., Correia, J.J., Xing, J., Rener, B. and Cheung, H.C. Structural studies of kinesin-nucleotide intermediates. J. Biol. Chem. 271:30121-30221, 1996.

Lobert, S., Vulevic, B. and Correia, J.J. Interaction of vinca alkaloids with tubulin: A comparison of vinblastine, vincristine and vinorelbine. Biochemistry 35:6806-6814, 1996. [link to PDF]

Lobert, S., Boyd, C.A. and Correia, J.J. Divalent cation and ionic strength effects in vinca alkaloid-induced tubulin self-association. Biophys. J. 72: 416-427, 1997.

Vulevic, B. and Correia, J.J. Thermodynamic and structural analysis of microtubule assembly: The role of GTP hydrolysis. Biophys. J. 72: 1357-1375, 1997.

Vulevic, B., Lobert, S. and Correia, J.J. Role of Guanine Nucleotides in the Vinblastine-Induced Self-Association of Tubulin: Effects of GMPCPP and GMPCP. Biochemistry 36: 12828-12835, 1997. [link to PDF]

Lobert, S., Frankfurter, A. and Correia, J.J. Energetics of Vinca Alkaloid Interactions with Tubulin Isotypes: Implications for Drug Efficacy and Toxicity. Cell Motility and Cytoskeleton 39: 107-121, 1998.

Lobert, S., Ingram, J.W., Hill, B.T. and Correia, J.J. A Comparison of Thermodynamic Parameters for Vinorelbine- and Vinflunine-Induced Tubulin Self Association by Sedimentation Velocity. Mol. Pharmacol. 53: 908-915, 1998.

Correia, J. J. Sedimentation velocity data analysis methods: what, when and why? CHEMTRACTS - Biochemistry and Molecular Biology, 11: 944-948, 1998. [link to Word file]

Foster, K. A., Correia, J. J. and Gilbert, S. P. Equilibrium Binding Studies of Non-claret Disjunctional Protein (Ncd). J. Biol. Chem. 273: 35307-35318, 1998.

Lobert, S., Ingram, J.W. and Correia, J.J. Additivity of Dilantin and Vinblastine Inhibitory Effects on Microtubule Assembly, Cancer Res., 59: 4816-4822, 1999.

Correia, J. J. The Analysis of Weight Average Sedimentation Data. Methods in Enzymology, Numerical Computer Methods, Part C, 321:81-100, 2000.

Lobert, S. and Correia, J.J. Energetics of Vinca Alkaloid Interactions with Tubulin. Methods in Enzymology, Energetics of Biological Macromolecules, Part C,323: 77-103, 2000.

Lobert, S., Fahy, J., Hill, B.T., Duflos, A., Etievant, C. and Correia, J.J. Vinca Alkaloid-Induced Tubulin Spiral Formation Correlates with Cytotoxicity in the Leukemic L1210 Cell Line. Biochemistry 39: 12053-12062, 2000. [link to PDF]

Correia, J.J., Chacko, B.M., Lam, S.S. and Lin, K. Sedimentation Studies Reveal a Direct Role of Phosphorylation in Smad3:Smad4 Homo- and Hetero-Trimerization. Biochemistry, 40: 1473-1482, 2001. [link to PDF]

Correia, J.J.and Lobert, S. Physiochemical Aspects of Tubulin-Interacting Antimitotic Drugs. Current Pharmaceutical Design,7(13): 1213-28, 2001. [link to PDF]

De Marco, V., de Marco, A., Correia, J.J., Goldie, K.N. and Hoenger, A. Dimerization Properties of a Xenopus Laevis Kinesin-II C-terminal Stalk Fragment. EMBO Reports 4: 717-722, 2003. [link to PDF]

J. Dozier, L. Hiser, J. A. Davis, N. Stubbs, S. Tucci, H. Bengouzzi, Anthony Frankfurter J. J. Correia and S.Lobert beta-class II Tubulin Predominates in Normal and Tumor Breast Tissues. Breast Cancer Res., 5: R157-R169, 2003. [link to PDF]

J. J. CORREIA, M. L. Johnson, T. Laue, W. F. Stafford and R. C. Williams, Jr., Spinning With Dave: David Yphantis's Contributions to Ultracentrifugation. Biophys. Chem. 108: 23-42, 2004. [link to PDF from Festschrift for Dave Yphantis]

Sontag,C.A., Stafford,W.F. and CORREIA, J.J. A Comparison of Weight Average and Direct Boundary Fitting of Sedimentation Velocity Data for Indefinite Polymerizing Systems. Biophys. Chem. 108: 215-230, 2004. [link to PDF from Festschrift for Dave Yphantis]

J. J. CORREIA, C. A. Sontag, W. F. Stafford and P. J. Sherwood, (2005) “Models for Direct Boundary Fitting of Indefinite Ligand-Linked Self-Association,”  in Analytical Ultracentrifugation: Techniques and Methods. (Ed. D. Scott, S. Harding and A. Rowe) pp 51-63.

L. Hiser, A. Aggarwal, R. Young, A. Frankfurter, A, Spano, J. J. CORREIA, and S. Lobert. "Correlation of Beta-Tubulin mRNA and Protein Levels in Twelve Human Cancer Cell Lines", Cell Motility & Cytoskeleton, 63: 41-52, 2006. [link to PDF]

S. Lobert and J.J. Correia (2006) "Methods for studying vinca alkaloid interaction with tubulin." Humana Press, (Ed., Jin Zhou), in press.

S. Lobert, J.W. Ingram and J.J. Correia (2007) "The Thermodynamics of Vinca Alkaloid-Induced Tubulin Spiral Formation." Biophys. Chem, Special Issue, Festschrift for Julian Sturtevant, 126: 50-58.

 Anthony J. Spano, Frank S. Chen, Benjamin E. Goodman, Agnes E. Sabat, Martha N. Simon, Joseph S. Wall,  John J. CORREIA, Wilson McIvor, William W. Newcomb, Jay C. Brown, Joel M. Schnur and Nikolai Lebedev.  (2007)  “In vitro assembly of a Prohead-like Structure of Rhodobacter capsulatus Gene Transfer Agent.”  Virology, 364: 95-102.

S. S. Dignam, J. J. CORREIA, S. A. Nada, J. P.  Trempe, J. D. Dignam (2007)  “Activation of the ATPase Activity of Adenoassociated Virus Rep68 and Rep78,” Biochemistry, 46: 6364-6374.

Montserrat Bosch, Kim Ho Diep Le, Beata Bugyi, John J. Correia, Louis Renault and Marie-France Carlier, (2007)  “Analysis of the function of Spire in actin assembly and its synergy with formin and profiline”, Mol Biol Cell, in press.

Ashish Aggarwal, Anna Kruczynski, Anthony Frankfurter, John J. Correia and Sharon Lobert, (2008) "Murine Leukemia P388 Vinorelbine-Resistant Cell Lines Are Sensitive to Vinflunine", Investigational New Drugs, in press.

M.A. Jordan, S.B. Horowitz, S. Lobert, J.J. Correia, (2008) "Exploring the Mechanisms of Action of the Novel Microtubule Inhibitor Vinflunine."  Seminars in Oncology Vol xx, S1-S7.

Valeria Cucchiarelli, Laree Hiser, Hilda Smith, Anthony Frankfurter, Anthony Spano, John J. Correia and Sharon Lobert , (2008) "β-Tubulin Isotype Classes II and V Expression Patterns in Nonsmall Cell Lung Carcinomas." Cell Motil. Cytoskel., in press.

J. J. CORREIA and S. Lobert (2008) "Molecular Mechanisms of Micotubule Acting Cancer Drugs," in Microtubules in Health and Disease, Humana Press (Ed., Tito Fojo), in press.