Citation

  • Authors: Andreu-Carbó M. et al.
  • Year: 2022
  • Journal: Dev Cell 57 5-18.e8
  • Applications: in vitro / DNA / jetOPTIMUS
  • Cell types:
    1. Name: HeLa
      Description: Human cervix epitheloid carcinoma cells
    2. Name: PtK2
      Description: Marsupial cell line.
      Known as:
      Pt K2 (NBL-5); NBL-5; Pt-K2; PTK-2; Ptk-2; PTK 2; PtK 2; PTK2; Ptk2; Potorous tridactylus Kidney 2.

Method

GFP-Tubulin Ptk2 and Hela cells were transfected with jetOPTIMUS transfection reagent (Polyplus) using 1 μg of plasmid in 24-well plates according to the manufacturer’s instructions. Cells were analyzed 8-24 hours after transfection.

Abstract

Tubulin dimers assemble into dynamic microtubules, which are used by molecular motors as tracks for intracellular transport. Organization and dynamics of the microtubule network are commonly thought to be regulated at the polymer ends, where tubulin dimers can be added or removed. Here, we show that molecular motors running on microtubules cause exchange of dimers along the shaft in vitro and in cells. These sites of dimer exchange act as rescue sites where depolymerizing microtubules stop shrinking and start re-growing. Consequently, the average length of microtubules increases depending on how frequently they are used as motor tracks. An increase of motor activity densifies the cellular microtubule network and enhances cell polarity. Running motors leave marks in the shaft, serving as traces of microtubule usage to organize the polarity landscape of the cell.

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