title: Coordinated interaction of Down syndrome cell adhesion molecule and deleted in colorectal cancer with dynamic TUBB3 mediates Netrin-1-induced axon branching.
abstract: Modulation of actin and microtubule (MT) dynamics in neurons is implicated in guidance cue-dependent axon outgrowth, branching and pathfinding. Although the role of MTs in axon guidance has been well known, how extracellular guidance signals engage MT behavior in axon branching remains unclear. Previously, we have shown that TUBB3, the most dynamic β-tubulin isoform in neurons, directly binds to deleted in colorectal cancer (DCC) to regulate MT dynamics in Netrin-1-mediated axon guidance. Here, we report that TUBB3 directly interacted with another Netrin-1 receptor Down syndrome cell adhesion molecule (DSCAM) and Netrin-1 increased this interaction in primary neurons. MT dynamics were required for Netrin-1-promoted association of DSCAM with TUBB3. Knockdown of either DSCAM or DCC or addition of a function blocking anti-DCC antibody mutually blocked Netrin-1-induced interactions, suggesting that DSCAM interdependently coordinated with DCC in Netrin-1-induced binding to TUBB3. Both DSCAM and DCC were partially colocalized with TUBB3 in the axon branch and the axon branching point of primary neurons and Netrin-1 increased these colocalizations. Netrin-1 induced the interaction of endogenous DSCAM with polymerized TUBB3 in primary neurons and Src family kinases (SFKs) were required for regulating this binding. Knockdown of DSCAM only, DCC only or both was sufficient to block Netrin-1-induced axon branching of E15 mouse cortical neurons. Knocking down TUBB3 inhibited Netrin-1 induced axon branching as well. These results suggest that DSCAM collaborates with DCC to regulate MT dynamics via direct binding to dynamic TUBB3 in Netrin-1-induced axon branching.
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA.
authors.forename: H
authors.initials: H
authors.lastname: Huang
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA.
authors.forename: Q
authors.initials: Q
authors.lastname: Shao
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA.
authors.forename: C
authors.initials: C
authors.lastname: Qu
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA.
authors.forename: T
authors.initials: T
authors.lastname: Yang
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA.
authors.forename: T
authors.initials: T
authors.lastname: Dwyer
authors.affiliation: Department of Biological Sciences, University of Toledo, 2801 West Bancroft Street, Toledo, OH 43606, USA. Electronic address: Guofa.Liu@utoledo.edu.
authors.forename: G
authors.initials: G
authors.lastname: Liu
mesh_terms: D000818:Animals; D001369:Axons; D015815:Cell Adhesion Molecules / Q000378:metabolism*; D002478:Cells, Cultured; D000075389:DCC Receptor; D020439:Growth Cones / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D051379:Mice; D008870:Microtubules / Q000378:metabolism; D009414:Nerve Growth Factors / Q000378:metabolism*; D000075388:Netrin-1; D011956:Receptors, Cell Surface / Q000378:metabolism*; D014404:Tubulin / Q000378:metabolism*; D025521:Tumor Suppressor Proteins / Q000378:metabolism*
chemical_list: D015815:Cell Adhesion Molecules; D000075389:DCC Receptor; C501910:Dcc protein, mouse; C491735:Dscam protein, mouse; C000617351:NTN1 protein, human; D009414:Nerve Growth Factors; C000617353:Ntn1 protein, mouse; D011956:Receptors, Cell Surface; C000592703:Tubb3 protein, mouse; D014404:Tubulin; D025521:Tumor Suppressor Proteins; D000075388:Netrin-1
keywords: DCC; DSCAM; Netrin-1; TUBB3; axon branching; microtubule dynamics