Pulses of Ca2+ coordinate actin assembly and exocytosis for stepwise cell extension

Takeshita Norio takeshita.norio.gf@u.tsukuba.ac.jp ^1,2 Minoas Evangelionos ² Lu Zhou ³ Tomato Serizawa ¹ Ling Lu ⁴ Naoki Takaya ¹ Ulrich Nienhaus ³ Reinhard Fischer ²

¹Faculty of Life and Environmental Sciences, University of Tsukuba, Tsukuba, Japan
²Department of Microbiology, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany
³Institute of Applied Physics, Karlsruhe Institute of Technology (KIT), Karlsruhe, Germany
⁴College of Life Sciences, Nanjing Normal University, Nanjing, China

Many eukaryotic cells grow by extending their cell periphery in pulses. The molecular mechanisms underlying this process are not yet fully understood. Here we present a comprehensive model of stepwise cell extension by using the unique tip growth system of filamentous fungi. Live-cell imaging analysis, including superresolution microscopy, revealed that the fungus Aspergillus nidulans extends the hyphal tip in an oscillatory manner. The amount of F-actin and secretory vesicles (SV) accumulating at the hyphal tip oscillated with a positive temporal correlation, whereas vesicle amounts were negatively correlated to the growth rate. The intracellular Ca2+ level also pulsed with a positive temporal correlation to the amount of F-actin and SV at the hyphal tip. Two Ca2+ channels, MidA and CchA, were needed for proper tip growth and the oscillations of actin polymerization, exocytosis, and the growth rate. The data indicate a model in which transient Ca2+ pluses cause depolymerization of F-actin at the cortex and promote SV fusion with the plasma membrane, thereby extending the cell tip. Over time, Ca2+ diffuses away and F-actin and SV accumulate again at the hyphal tip. Our data provide evidence that temporally controlled actin polymerization and exocytosis are coordinated by pulsed Ca2+ influx, resulting in stepwise cell extension.

Takeshita et al, PNAS, 114(22):5710-06, 2017.