Subcellular localization of S100A11 (S100C) in LLC-PK1 renal cells: Calcium- and protein kinase c-dependent association of S100A11 with S100B and vimentin intermediate filaments.
The subcellular localization of the Ca(2+)-modulated protein, S100A11, was investigated in the renal cell line LLC-PK1 by immunofluorescence and confocal laser scanning microscopy under varying experimental conditions. In control cells, S100A11 was detected on the plasma membrane, where the protein co-localized with annexin I (ANXA1) at discrete sites, and found diffusely in the cytoplasm. Elevation of the cytosolic Ca(2+) concentration by means of the Ca(2+) ionophore, ionomycin, caused a significant fraction of S100A11 to associate with vimentin intermediate filament (IF)-bound S100B, another member of the S100 protein family. Under these conditions, ANXA1 underwent a quite different kind of relocation. Translocation of S100A11 onto vimentin IF-bound S100B was also observed upon activation of protein kinase C (PKC). Under these conditions, S100A11 appeared to associate directly with vimentin IFs at cell sites displaying low or no abundance of S100B such as cell processes, and, again, S100A11 and ANXA1 underwent a different relocation. Our data suggest the possibility that the intracellular Ca(2+) level might regulate the subcellular localization of S100A11 and its interaction with definite target proteins, and that S100A11 might serve the function of modulating S100B activities. Interestingly, in spite of the known ability of S100A11 to form heterotetramers with ANXA1, the two proteins underwent a different relocation on elevation of the cytosolic Ca(2+) concentration or activation of PKC, pointing to different regulatory activities of individual proteins in renal cells.
Bianchi
Roberta
R
Department of Experimental Medicine and Biochemical Sciences, Section of Anatomy, University of Perugia, 06122 Perugia, Italy.
Giambanco
Ileana
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Arcuri
Cataldo
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Donato
Rosario
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D000818:Animals; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D005455:Fluorescent Antibody Technique; D007382:Intermediate Filaments / Q000378:metabolism*; D018374:LLC-PK1 Cells / Q000378:metabolism* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D009414:Nerve Growth Factors / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism*; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins / Q000378:metabolism*; D013552:Swine; D014746:Vimentin / Q000378:metabolism
D009414:Nerve Growth Factors; D064568:S100 Calcium Binding Protein beta Subunit; D009418:S100 Proteins; D014746:Vimentin; D011493:Protein Kinase C; D002118:Calcium