Selective effects of calcium chelators on anterograde and retrograde protein transport in the cell.
Calcium plays a regulatory role in several aspects of protein trafficking in the cell. Both vesicle fusion and vesicle formation can be inhibited by the addition of calcium chelators. Because the effects of calcium chelators have been studied predominantly in cell-free systems, it is not clear exactly which transport steps in the secretory pathway are sensitive to calcium levels. In this regard, we have studied the effects of calcium chelators on both anterograde and retrograde protein transport in whole cells. Using both cytochemical and biochemical analyses, we find that the anterograde-directed exit of vesicular stomatitis virus G protein and the retrograde-directed exit of Shiga toxin from the Golgi apparatus are both inhibited by calcium chelation. The exit of vesicular stomatitis virus G from a pre-Golgi compartment and the exit of Shiga toxin from an endosomal compartment are sensitive to the membrane-permeant calcium chelator 1,2-bis(2-amino phenoxy)ethane-N,N,N',N'-tetraacetic acid-tetrakis (acetoxymethyl ester) (BAPTA-AM). By contrast, endoplasmic reticulum exit and endocytic internalization from the plasma membrane are not affected by BAPTA. Together, our data show that some, but not all, trafficking steps in the cell may be regulated by calcium. These studies provide a framework for a more detailed analysis of the role of calcium as a regulatory agent during protein transport.
Department of Physiology & Biophysics, Roy J. and Lucille A. Carver College of Medicine, The University of Iowa, Iowa City, Iowa 52242, USA.
Ji-Long
JL
Chen
Jatinder P
JP
Ahluwalia
Mark
M
Stamnes
D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002614:Chelating Agents / Q000378:metabolism*; D017931:DNA Primers; D004533:Egtazic Acid / Q000031:analogs & derivatives* / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D008562:Membrane Glycoproteins; D021381:Protein Transport; D014759:Viral Envelope Proteins / Q000378:metabolism
D002614:Chelating Agents; D017931:DNA Primers; C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D004533:Egtazic Acid; D002118:Calcium