Phospholipase D-dependent and -independent mechanisms are involved in milk protein secretion in rabbit mammary epithelial cells.
Phospholipase D has been implicated in membrane traffic in the secretory pathway of yeast and of some mammalian cell lines. Here we investigated the involvement of phospholipase D in protein transport at various steps of the secretory pathway of mammary epithelial cells. Treatment of rabbit mammary explants with butanol, which blocks the formation of phosphatidic acid, decreased the secretion of caseins and to a lesser extent that of whey acidic protein. Butanol interfered with both the endoplasmic reticulum to Golgi complex transport of the caseins and secretory vesicle formation from the trans-Golgi network. In contrast, the transport of whey acidic protein to the Golgi was less affected. Activation of protein kinase C enhanced the overall secretion of both markers and interestingly, this stimulation of secretion was maintained for whey acidic protein in the presence of butanol. Transphosphatidylation assays demonstrated the existence of a constitutive phospholipase D activity which was stimulated by the activation of protein kinase C. We conclude that phospholipase D plays a role in casein transport from the endoplasmic reticulum to the Golgi and in the secretory vesicle formation from the trans-Golgi network. Moreover, our results suggest a differential requirement for phospholipase D in the secretion of caseins and that of whey acidic protein.
Boisgard
R
R
Institut National de la Recherche Agronomique, Laboratoire de Biologie Cellulaire et Moleculaire, F-78352, Jouy-en-Josas, France.
Chanat
E
E
D000818:Animals; D001692:Biological Transport; D001940:Breast / Q000201:enzymology* / Q000378:metabolism; D000440:Butanols / Q000494:pharmacology; D002364:Caseins / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004847:Epithelial Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D005260:Female; D006056:Golgi Apparatus / Q000378:metabolism; D007700:Kinetics; D008894:Milk Proteins / Q000378:metabolism*; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology; D010739:Phospholipase D / Q000502:physiology*; D011817:Rabbits; D012741:Sexual Maturation / Q000502:physiology; D012799:Sialyltransferases / Q000378:metabolism; D013347:Subcellular Fractions / Q000378:metabolism
D000440:Butanols; D002364:Caseins; D008894:Milk Proteins; C024471:whey acidic proteins; D015240:Phorbol 12,13-Dibutyrate; D012799:Sialyltransferases; D010739:Phospholipase D