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With the emergences of engineered devices at microscale and nanoscale dimensions, there is a growing need for controlled actuation and transport at these length scales. The kinesin-microtubule system provides a highly evolved biological transport system well suited for these tasks. Accordingly, there is an ongoing effort to create hybrid nanodevices that integrate biological components with engineered materials for applications such as biological separations, nanoscale assembly, and sensing. Adopting microtubules for these applications generally requires covalent attachment of biotin, fluorophores, or other biomolecules to tubulin enable surface or cargo attachment, or visualization. This review summarizes different strategies for functionalizing microtubules for application-focused as well as basic biological research. These functionalization strategies must maintain the integrity of microtubule proteins so that they do not depolymerize and can be transported by kinesin motors, while adding utility such as the ability to reversibly bind cargo. The relevant biochemical and electrical properties of microtubules are discussed, as well as strategies for microtubule stabilization and long-term storage. Next, attachment strategies, such as antibodies and DNA hybridization that have proven useful to date, are discussed in the context of ongoing hybrid nanodevice research. The review concludes with a discussion of less explored opportunities, such as harnessing the utility of tubulin posttranslational modifications and the use of recombinant tubulin that may enable future progress in nanodevice development.

Past phylogenic studies have identified a plant-specific, ungrouped family of kinesins in which the motor domain does not group to one of the fourteen recognized families. Members of this family contain an N-terminal motor domain, a C-terminal armadillo repeat domain and a conserved destruction box (D-BOX) motif. This domain architecture is unique to plants and to a subset of protists. Further characterization of one representative member from Arabidopsis, Arabidopsis thaliana KINESIN ungrouped clade, gene A (AtKINUa), was completed to ascertain its functional role in plants. Fluorescence confocal microscopy revealed an accumulation of ATKINUA:GFP at the preprophase band (PPB) in a cell cycle-dependent manner in Arabidopsis epidermal cells and tobacco BY-2 cells. Fluorescence accumulation was highest during prophase and decreased after nuclear envelope breakdown. A conserved D-BOX motif was identified through alignment of AtKINU homologous sequences. Mutagenesis work with D-BOX revealed that conserved residues were necessary for the observed degradation pattern of ATKINUA:GFP, as well as the targeted accumulation at the PPB. Overall results suggest that AtKINUa is necessary for normal plant growth and/or development and is likely involved with PPB organization through microtubule association and specific cell cycle regulation. The D-BOX motif may function to bridge microtubule organization with changes that occur during progression through mitosis and may represent a novel regulatory motif in plant microtubule motor proteins., (Copyright © 2011 Wiley-Liss, Inc.)

Gene duplication is important for gene family evolution, allowing for functional divergence and innovation. In flowering plants, duplicated genes are widely observed, and functional redundancy of closely related duplicates has been reported, but few cases of functional divergence of close duplicates have been described. Here, we show that the Arabidopsis AtKIN14a and AtKIN14b genes encoding highly similar kinesins are two of the most closely related Arabidopsis paralogs, which were formed by a duplication event that occurred after the split of Arabidopsis and poplar. In addition, AtKIN14a and AtKIN14b exhibit varying degrees of coding sequence divergence. Further genetic studies of plants carrying atkin14a and/or atkin14b mutations indicate that, although these two genes have similar functions, there is clear evidence for functional divergence. Although both genes are important for male and female meiosis, AtKIN14a plays a more critical role in male meiosis than AtKIN14b. Moreover, either one of these two genes is necessary and sufficient for gametophyte development, indicating that they are redundant for this function. Therefore, AtKIN14a and AtKIN14b together play important roles in controlling plant reproductive development. Our results suggest that the AtKIN14a and AtKIN14b genes have retained similar functions in gametophyte development and female meiosis, but have evolved partially distinct functions in male meiosis, with AtKIN14a playing a more substantive role.