Spindle assembly is the crucial initiating step in the mitotic mechanism. Its formation utilizes coordinated actions of Kinesin-like proteins (Klps) at poles, on microtubules and at chromosomes. We previously identified a novel conserved Kinesin-14 pole-based mechanism for spindle assembly by Klp binding to the γ-tubulin small complex (γ-TuSC) microtubule organizing center, (MTOC). Changes to the γ-TuSC MTOC in fission yeast alter microtubule dynamics to regulate bipolarity and both human and fission yeast Kinesin-14 Klps can regulate this mechanism. The Kinesin-14 Pkl1 motor domain associates with residues in helix 11 of γ-tubulin at a novel Klp binding site. The Kinesin-14 Pkl1 Tail domain is distinct from that of the well-characterized Drosophila Ncd, replacing microtubule association sites with specialized elements for spindle pole targeting and regulation of the γ-TuSC. Thus unlike Drosophila Ncd that crosslinks microtubules to stabilize mitotic spindle assembly, Kinesin-14 Pkl1 regulates assembly through the γ-TuSC. Klps can be classified into 14 families. Members of two ubiquitous families, Kinesin-14 and opposing Kinesin-5 Klps, each exhibit distinctions at multiple levels amongst members, including domain elements, functional mechanisms and spindle localization. In fission yeast, Kinesin-5 also localizes to spindle poles, consistent with the hypothesis that similar localization is needed to optimally counterbalance forces by these two Klp families. Although the stoichiometry of proteins within γ-TuSC is known and conserved in eukaryotes little is understood of the γ-TuSC mechanism. Cross-species analysis is defining shared and distinct functions and novel insights into the γ-TuSC interactome and the role of associating Klps in regulating its mechanism for spindle assembly. Our studies apply site directed mutagenesis, genetics, yeast two hybrid, biochemistry, timelapse microscopy and bioinformatic/structural approaches with novel reagents we have developed for Klp and γ-TuSC analysis.