title: Insights into antiparallel microtubule crosslinking by PRC1, a conserved nonmotor microtubule binding protein.
abstract: Formation of microtubule architectures, required for cell shape maintenance in yeast, directional cell expansion in plants and cytokinesis in eukaryotes, depends on antiparallel microtubule crosslinking by the conserved MAP65 protein family. Here, we combine structural and single molecule fluorescence methods to examine how PRC1, the human MAP65, crosslinks antiparallel microtubules. We find that PRC1's microtubule binding is mediated by a structured domain with a spectrin-fold and an unstructured Lys/Arg-rich domain. These two domains, at each end of a homodimer, are connected by a linkage that is flexible on single microtubules, but forms well-defined crossbridges between antiparallel filaments. Further, we show that PRC1 crosslinks are compliant and do not substantially resist filament sliding by motor proteins in vitro. Together, our data show how MAP65s, by combining structural flexibility and rigidity, tune microtubule associations to establish crosslinks that selectively "mark" antiparallel overlap in dynamic cytoskeletal networks.
authors.affiliation: Laboratory of Chemistry and Cell Biology, The Rockefeller University, New York, NY 10065, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Subramanian
authors.forename: Elizabeth M
authors.initials: EM
authors.lastname: Wilson-Kubalek
authors.forename: Christopher P
authors.initials: CP
authors.lastname: Arthur
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Bick
authors.forename: Elizabeth A
authors.initials: EA
authors.lastname: Campbell
authors.forename: Seth A
authors.initials: SA
authors.lastname: Darst
authors.forename: Ronald A
authors.initials: RA
authors.lastname: Milligan
authors.forename: Tarun M
authors.initials: TM
authors.lastname: Kapoor
mesh_terms: D018797:Cell Cycle Proteins / Q000737:chemistry / Q000378:metabolism*; D020285:Cryoelectron Microscopy; D006801:Humans; D008870:Microtubules / Q000378:metabolism*; D008958:Models, Molecular; D017434:Protein Structure, Tertiary; D013049:Spectrin / Q000378:metabolism
chemical_list: D018797:Cell Cycle Proteins; C116662:PRC1 protein, human; D013049:Spectrin