This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The objective of this research project is to visualize the interactions between Kinesin-13 and tubulin rings. The kinesin family of motor proteins utilize energy derived from ATP-hydrolysis to bind, and do work, on microtubules (Vale and Fletterick, 1997. Annu. Rev. Cell Dev. Biol. 13: 745-777). The dynamic nature of microtubules is important for fundamental cellular processes such as spindle assembly during mitosis, cell migration, and cellular transport;in fact, it is the interactions with microtubule associated proteins (MAPS), motor proteins such as kinesin, and other cellular factors, that regulate the dynamic behavior, and so the function, of microtubules (Walczack, 2000. Curr. Opin. Cell. Biol. 12: 52-56). Kinesins are most known for their ability to walk along a microtubular track while carrying cargo from one point in the cell to another;however the intermediate kinesins, of which Kinesin-13 is a member, have not been shown to engage in such a walking motion. Instead, this family of kinesins utilize chemical energy to depolymerize the ends of microtubules. Previous studies have shown that, in the process of depolymerizing microtubule ends, tubulin rings and spirals are formed (Moores et. al. 2002. Mol. Cell. 9: 903-909;Desai et. al. 1999 Cell. 96: 69-78);importantly, addition of Dolastatin to alpha and beta tubulin results in the spontaneous formation of rings indistinguisable from those formed during the depolymerization process (unpublished results). It is thought that these tubulin rings are representative of tubulin configuration at the ends of microtubules, and that studies of motors bound to such rings will give important insights regarding the molecular mechanism of microtubule depolymerization. In the first phase of this project, we will utilize the motor domain of KinI from Plasmodium falciparum (pKinI) to institute an automated platform for data collection and processing via LEGINON and single particle 2D averaging techniques. Once, an automated platform has been established, we will extend our studies to include full-length human kinesin-13 and various truncated versions of the protein. It is the hope that such studies will contribute to our molecular understanding of microtubule depolymerization.
| Scapin, Giovanna; Dandey, Venkata P; Zhang, Zhening et al. (2018) Structure of the insulin receptor-insulin complex by single-particle cryo-EM analysis. Nature 556:122-125 |
| Sherman, Michael B; Kakani, Kishore; Rochon, D'Ann et al. (2017) Stability of Cucumber Necrosis Virus at the Quasi-6-Fold Axis Affects Zoospore Transmission. J Virol 91: |
| Geary, Cody; Chworos, Arkadiusz; Verzemnieks, Erik et al. (2017) Composing RNA Nanostructures from a Syntax of RNA Structural Modules. Nano Lett 17:7095-7101 |
| Kulczyk, Arkadiusz W; Moeller, Arne; Meyer, Peter et al. (2017) Cryo-EM structure of the replisome reveals multiple interactions coordinating DNA synthesis. Proc Natl Acad Sci U S A 114:E1848-E1856 |
| Razinkov, Ivan; Dandey, Venkat; Wei, Hui et al. (2016) A new method for vitrifying samples for cryoEM. J Struct Biol 195:190-198 |
| Short, James R; Speir, Jeffrey A; Gopal, Radhika et al. (2016) Role of Mitochondrial Membrane Spherules in Flock House Virus Replication. J Virol 90:3676-83 |
| Guenaga, Javier; de Val, Natalia; Tran, Karen et al. (2015) Well-ordered trimeric HIV-1 subtype B and C soluble spike mimetics generated by negative selection display native-like properties. PLoS Pathog 11:e1004570 |
| McCullough, John; Clippinger, Amy K; Talledge, Nathaniel et al. (2015) Structure and membrane remodeling activity of ESCRT-III helical polymers. Science 350:1548-51 |
| McNulty, Reginald; Lokareddy, Ravi Kumar; Roy, Ankoor et al. (2015) Architecture of the Complex Formed by Large and Small Terminase Subunits from Bacteriophage P22. J Mol Biol 427:3285-3299 |
| Lee, Jeong Hyun; Leaman, Daniel P; Kim, Arthur S et al. (2015) Antibodies to a conformational epitope on gp41 neutralize HIV-1 by destabilizing the Env spike. Nat Commun 6:8167 |
Showing the most recent 10 out of 189 publications
