The role of protein kinase C in the transient association of p57, a coronin family actin-binding protein, with phagosomes.
Phagocytosis of opsonized zymosan (OpZ) particles by differentiated cells of the human leukemic cell line HL-60 induced transient periphagosomal association of p57, a coronin family actin-binding protein, and F-actin with dissociation from the phagosomes after ingestion was completed. Coincident with OpZ ingestion, p57 phosphorylation increased transiently and peaked with its dissociation from phagosomes. Since p57 contains several putative sites for protein kinase C (PKC) phosphorylation, we examined the effect of PKC on p57 phosphorylation and association with the phagosome. Purified p57 was phosphorylated in vitro by PKC isoforms alpha and delta, and PMA, an activator of PKC, induced p57 phosphorylation in HL-60 cells. Furthermore, chelerythrine, a specific PKC inhibitor, blocked p57 phosphorylation and the dissociation of p57 and F-actin from phagosomes, whereas wortmannin, genistein, and H-89 did not. Chelerythrine also inhibited the translocation of LAMP-1, a marker protein of lysosomes, to the OpZ-containing phagosomes, indicating that PKC-mediated phosphorylation is required for phagosome-lysosome fusion. Taken together, these data suggest that PKC-mediated phosphorylation of p57 triggers its dissociation from phagosomes, an event that may be necessary for the fusion of phagosomes with lysosomes.
Department of Biochemistry, Hoshi University, Tokyo, Japan. s-itoh@hoshi.ac.jp
Saotomo
S
Itoh
Kensuke
K
Suzuki
Jun
J
Nishihata
Mitsusada
M
Iwasa
Teruaki
T
Oku
Shizuo
S
Nakajin
William Michael
WM
Nauseef
Satoshi
S
Toyoshima
D000199:Actins / Q000378:metabolism*; D002352:Carrier Proteins; D018922:HL-60 Cells; D006801:Humans; D008840:Microfilament Proteins / Q000378:metabolism*; D008954:Models, Biological; D010587:Phagocytosis / Q000502:physiology*; D010588:Phagosomes / Q000378:metabolism*; D010766:Phosphorylation; D011485:Protein Binding; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000502:physiology*; D015054:Zymosan / Q000494:pharmacology
D000199:Actins; D002352:Carrier Proteins; C034037:F-actin-binding proteins; D008840:Microfilament Proteins; D015054:Zymosan; D011493:Protein Kinase C