Structural and Functional Characterization of the Regulation Network of Microtubule Plus-ends using Hybrid Methods

• Applicant: Dr. Naoko Mizuno
• Subject Area: Structural Biology
• Term: from 2014 to 2018
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 254667228

• Final Report Year: 2018

Final Report Abstract

p150 is a member of +TIPs, accumulating at microtubule plus ends. It plays a key role in initiating retrograde transport by recruiting and dynein. p150 contains a microtubule-binding CAP-Gly domain that accelerates tubulin polymerization. While this copolymerization is well studied using light microscopic techniques, structural consequences of the interaction are elusive. In this proposed research, we published a research article presenting a direct visualization of the microtubule-CAP-Gly by cryo-EM and showed the molecular mechanism of the control of microtubule dynamics. The neutralization of acidic tubulin surface by the basic extension of CAP-Gly resulted in the activation of the lateral association. In the condition where the lateral association is impeded, the extended CAP-Gly induces the longitudinal tubulin association. These two directional modes of self-associations induced by CAP-Gly may provide a foundation for the microtubule dynamic behavior.

Publications

• (2014) Architecture and ssDNA interaction of the Timeless-Tipin- RPA Complexs. Nucleic Acids Research 42:12912-27
  Witosch J, Wolf E, and Mizuno N
  
• (2014) Cofilin recruits F-actin to SPCA1 and promotes Ca2+-mediated secretory cargo sorting. J Cell Biol 206:635-54
  Kienzle C, Basnet N, Crevenna AH, Beck G, Habermann B, Mizuno N, von Blume J
  
• (2014) Structural basis for the extended CAP-Gly domains of p150(glued) binding to microtubules and the implication for tubulin dynamics. Proc Natl Acad Sci USA. 111:11347-52
  Wang Q, Crevenna AH, Kunze I and Mizuno N
  
• (2015) Side-binding proteins modulate actin filament dynamics. Elife. Feb 23;4
  Crevenna AH, Arciniega M, Dupont A, Mizuno N, Kowalska K, Lange OF, Wedlich-Söldner R, Lamb DC
  
• (2015) Structural insights into the cooperative remodeling of membranes by amphiphysin/BIN1. Sci Rep. Oct 21;5:15452
  Adam J, Basnet N, Mizuno N
  
• (2016) Kank2 activates Talin and reduce force transduction across integrins. Nature Cell Biol. 18:941-53
  Sun Z, Tseng H, Tan S, Senger F, Kurzawa L, Dedden D, Mizuno N, Wasik A, Thery M, Dunn A, Fässler R