Calcium-dependent translocation of S100A11 requires tubulin filaments.
Protein translocation between different subcellular compartments might play a significant role in various signal transduction pathways. The S100 family is comprised of the multifunctional, small, acidic proteins, some of which translocate in the form of vesicle-like structures upon increase in intracellular Ca(2+) levels. Previously, cells were fixed before and after calcium activation in order to examine the possible relocation of S100 proteins. In this study, we were able to track the real-time translocation. We compared the localization of endogenous S100A11 to that of the S100A11-green fluorescent protein. The application of thapsigargin, an agent increasing intracellular Ca(2+) levels, resulted in the relocation of the S100A11. In contrast, addition of EGTA, which specifically binds Ca(2+), either inhibited the ongoing process of translocation or prevented its induction. Since translocation was not affected by treatment with brefeldin A, it appears that S100A11 relocates in an endoplasmic reticulum-Golgi-independent pathway. Furthermore, the depolymerization of actin filaments by amlexanox did not affect the capacity of S100A11 to translocate. However, the time course treatment with demecolcine, which depolymerizes tubulin filaments, resulted in cease of translocation, suggesting that the tubulin network is required for this process.
Davey
G E
GE
Department of Pediatrics, Division of Clinical Chemistry and Biochemistry, University of Zurich, Switzerland.
Murmann
P
P
Hoechli
M
M
Tanaka
T
T
Heizmann
C W
CW
D008841:Actin Cytoskeleton / Q000378:metabolism; D001692:Biological Transport; D002118:Calcium / Q000378:metabolism*; D004533:Egtazic Acid; D004721:Endoplasmic Reticulum / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D006056:Golgi Apparatus / Q000378:metabolism; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018613:Microscopy, Confocal; D009418:S100 Proteins / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D019284:Thapsigargin; D014162:Transfection; D014404:Tubulin / Q000378:metabolism*; D014407:Tumor Cells, Cultured
D008164:Luminescent Proteins; D009418:S100 Proteins; D014404:Tubulin; C072337:S100A11 protein, human; D049452:Green Fluorescent Proteins; D004533:Egtazic Acid; D019284:Thapsigargin; D002118:Calcium