Calmodulin regulates the disassembly of cortical F-actin in mast cells but is not required for secretion.
Secretion is dependent on a rise in cytosolic Ca(2+)concentration and is associated with dramatic changes in actin organization. The actin cortex may act as a barrier between secretory vesicles and plasma membrane. Thus, disassembly of this cortex should precede late steps of exocytosis. Here we investigate regulation of both the actin cytoskeleton and secretion by calmodulin. Ca(2+), together with ATP, induces cortical F-actin disassembly in permeabilized rat peritoneal mast cells. This effect is strongly inhibited by removing endogenous calmodulin (using calmodulin inhibitory peptides), and increased by exogenous calmodulin. Neither treatment, however, affects secretion. Low concentrations ( approximately 1 microM) of a specific inhibitor of myosin light chain kinase, ML-7, prevent F-actin disassembly, but not secretion. In contrast, a myosin inhibitor affecting both conventional and unconventional myosins, BDM, decreases cortical disassembly as well as secretion. Observations of fluorescein-calmodulin, introduced into permeabilized cells, confirmed a strong (Ca(2+)-independent) association of calmodulin with the actin cortex. In addition, fluorescein-calmodulin enters the nuclei in a Ca(2+)-dependent manner. In conclusion, calmodulin promotes myosin II-based contraction of the membrane cytoskeleton, which is a prerequisite for its disassembly. The late steps of exocytosis, however, require neither calmodulin nor cortical F-actin disassembly, but may be modulated by unconventional myosin(s).
Sullivan
R
R
Physiology Department, University College London, London, UK.
Burnham
M
M
Török
K
K
Koffer
A
A
D000199:Actins / Q000187:drug effects / Q000378:metabolism*; D021581:Active Transport, Cell Nucleus; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001381:Azepines / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000037:antagonists & inhibitors / Q000502:physiology*; D002148:Calmodulin-Binding Proteins / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003287:Contrast Media / Q000494:pharmacology; D003931:Diacetyl / Q000031:analogs & derivatives* / Q000494:pharmacology; D005089:Exocytosis / Q000187:drug effects; D019793:Fluorescein / Q000494:pharmacology; D008407:Mast Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D009219:Myosin-Light-Chain Kinase / Q000037:antagonists & inhibitors / Q000378:metabolism; D009218:Myosins / Q000037:antagonists & inhibitors; D009281:Naphthalenes / Q000494:pharmacology; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010455:Peptides / Q000378:metabolism; D051381:Rats
D000199:Actins; D001381:Azepines; D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D003287:Contrast Media; D009281:Naphthalenes; D010455:Peptides; C070571:ML 7; C004717:diacetylmonoxime; D000255:Adenosine Triphosphate; D019001:Nitric Oxide Synthase; D009219:Myosin-Light-Chain Kinase; D009218:Myosins; D003931:Diacetyl; D002118:Calcium; D019793:Fluorescein