title: Analysis of microtubule growth dynamics arising from altered actin network structure and contractility in breast tumor cells.
abstract: The periphery of epithelial cells is shaped by opposing cytoskeletal physical forces generated predominately by two dynamic force generating systems-growing microtubule ends push against the boundary from the cell center, and the actin cortex contracts the attached plasma membrane. Here we investigate how changes to the structure and dynamics of the actin cortex alter the dynamics of microtubules. Current drugs target actin polymerization and contraction to reduce cell division and invasiveness; however, the impacts on microtubule dynamics remain incompletely understood. Using human MCF-7 breast tumor cells expressing GFP-tagged microtubule end-binding-protein-1 (EB1) and coexpression of cytoplasmic fluorescent protein mCherry, we map the trajectories of growing microtubule ends and cytoplasmic boundary respectively. Based on EB1 tracks and cytoplasmic boundary outlines, we calculate the speed, distance from cytoplasmic boundary, and straightness of microtubule growth. Actin depolymerization with Latrunculin-A reduces EB1 growth speed as well as allows the trajectories to extend beyond the cytoplasmic boundary. Blebbistatin, a direct myosin-II inhibitor, reduced EB1 speed and yielded less straight EB1 trajectories. Inhibiting signaling upstream of myosin-II contractility via the Rho-kinase inhibitor, Y-27632, altered EB1 dynamics differently from Blebbistatin. These results indicate that reduced actin cortex integrity can induce distinct alterations in microtubule dynamics. Given recent findings that tumor stem cell characteristics are increased by drugs which reduce actin contractility or stabilize microtubules, it remains important to clearly define how cytoskeletal drugs alter the interactions between these two filament systems in tumor cells.
authors.affiliation: Department of Physics, IPST, and IREAP, University of Maryland, College Park, MD, United States of America.
authors.forename: Eleanor C
authors.initials: EC
authors.lastname: Ory
authors.forename: Lekhana
authors.initials: L
authors.lastname: Bhandary
authors.forename: Amanda E
authors.initials: AE
authors.lastname: Boggs
authors.forename: Kristi R
authors.initials: KR
authors.lastname: Chakrabarti
authors.forename: Joshua
authors.initials: J
authors.lastname: Parker
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Losert
authors.forename: Stuart S
authors.initials: SS
authors.lastname: Martin
mesh_terms: D008841:Actin Cytoskeleton / Q000378:metabolism*; D000199:Actins / Q000378:metabolism*; D006801:Humans; D008164:Luminescent Proteins / Q000378:metabolism; D061986:MCF-7 Cells; D008870:Microtubules / Q000378:metabolism*; D000097573:Red Fluorescent Protein
chemical_list: D000199:Actins; D008164:Luminescent Proteins; D000097573:Red Fluorescent Protein