Phorbol ester promotes endocytosis by activating a factor involved in endosome fusion

Alejandro Aballay, Philip D. Stahl, Luis S. Mayorga

Research output: Contribution to journalArticlepeer-review

22 Scopus citations

Abstract

Previous studies indicate that a zinc- and phorbol ester-binding factor is necessary for in vitro endosome fusion and for the effect of Rab5 on endosome fusion. Rab5 is a small GTPase that regulates membrane fusion between early endosomes derived from either receptor-mediated endocytosis or fluid-phase endocytosis. In its GTP-bound form, Rab5 promotes endocytosis and enhances fusion among early endosomes. To determine if PMA stimulates endocytosis by activating a factor required for endosome fusion, we overexpressed wild-type Rab5, a dominant negative mutant (Rab5:S34N), and a GTPase deficient mutant (Rab5:Q79L) in BHK-21 cells. The phorbol ester PMA stimulates endocytosis and increases the number and the size of endocytic vesicles, even in the presence of Rab5:S34N. Zinc depletion with N,N,N',N'-tetrakis-(2-pyridylmethyl)ethylenediamine (TPEN) and addition of calphostin C (CPC), an inhibitor of PKC that interacts with zinc and phorbol ester binding motifs, inhibited both basal and Rab5-stimulated fluid phase endocytosis. These two reagents also inhibited the size and number of endocytic vesicles promoted by Rab5. These results suggest that PMA stimulates endocytosis by regulating the dynamics of the early endosome compartment.

Original languageEnglish (US)
Pages (from-to)2549-2557
Number of pages9
JournalJournal of Cell Science
Volume112
Issue number15
StatePublished - Sep 2 1999

Keywords

  • PKC
  • PMA
  • Rab protein
  • Rab5
  • Zinc

ASJC Scopus subject areas

  • Cell Biology

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