Converging populations of f-actin promote breakage of associated microtubules to spatially regulate microtubule turnover in migrating cells.
BACKGROUND
In migrating cells, the retrograde flow of filamentous actin (f-actin) from the leading edge toward the cell body is accompanied by the synchronous motion of microtubules (MTs, ), whose plus ends undergo net growth. Thus, MTs must depolymerize elsewhere in the cell to maintain polymer mass over time. The source and location of depolymerized MTs is unknown. Here, we test the hypothesis that MT polymer loss occurs in central cell regions and is induced by the convergence of actin retrograde and anterograde flow, which buckles and breaks associated MTs and promotes minus-end depolymerization.


RESULTS
We characterized the effects of calyculin A and ML-7 on the movement of f-actin and MTs by multi-spectral fluorescence recovery after photobleaching (FRAP) and fluorescent speckle microscopy (FSM). Our studies show that these drugs affect the rate of f-actin and MT convergence and MT buckling in a central cell region we call the "convergence zone." Increases in f-actin convergence are associated with faster MT turnover and an increase in both MT breakage and minus-end depolymerization, but they have no effect on MT plus end dynamic instability.


CONCLUSIONS
We propose that f-actin movement into the convergence zone plays a major role in spatially modulating MT turnover during cell migration by regulating MT breakage, and thus minus-end dynamics, in central cell regions.
Gupton
Stephanie L
SL
Department of Cell Biology and Institute for Childhood and Neglected Diseases, The Scripps Research Institute, La Jolla, CA 92037, USA.
Salmon
Wendy C
WC
Waterman-Storer
Clare M
CM
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002465:Cell Movement / Q000502:physiology; D002478:Cells, Cultured; D007091:Image Processing, Computer-Assisted; D008168:Lung / Q000166:cytology / Q000502:physiology*; D008870:Microtubules / Q000502:physiology* / Q000648:ultrastructure*; D020545:Respiratory Mucosa / Q000502:physiology / Q000648:ultrastructure; D012455:Salamandridae
D000199:Actins