PKCζ is required for microtubule-based motility of vesicles containing the ntcp transporter

Souvik Sarkar, Eustratios Bananis, Sangeeta Nath, M. Sawkat Anwer, Allan W. Wolkoff, John W. Murray

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

Intracellular trafficking regulates the abundance and therefore activity of transporters present at the plasma membrane. The transporter, Na+-taurocholate co-transporting polypeptide (ntcp), is increased at the plasma membrane upon treatment of cells with cAMP, for which microtubules (MTs) are required and the PI3K pathway and PKCζ have been implicated. However, trafficking of ntcp on MTs has not been demonstrated directly and the regulation and intracellular localization of ntcp is not well understood. Here, we utilize in vitro and whole-cell immunofluorescence microscopy assays to demonstrate that ntcp is present on intracellular vesicles that bind MTs and move bidirectionally, using kinesin-1 and dynein. These vesicles co-localize with markers for recycling endosomes and early but not late endosomes. They frequently undergo fission, providing a mechanism for the exclusion of ntcp from late endosomes. PI(3,4,5)P3 activates PKCζ and enhances motility of the ntcp vesicles and overcomes the partial inhibition produced by a PI3-kinase inhibitor. Specific inhibition of PKCζ blocks the motility of ntcp-containing vesicles but has no effect on late vesicles as shown both in vitro and in living cells transfected with ntcp-GFP. These data indicate that PKCζ is required specifically for the intracellular movement of vesicles that contain the ntcp transporter.

Original languageEnglish (US)
Pages (from-to)1078-1091
Number of pages14
JournalTraffic
Volume7
Issue number8
DOIs
StatePublished - Aug 2006

Fingerprint

Taurocholic Acid
Microtubules
Peptides
Endosomes
Cell membranes
Phosphatidylinositol 3-Kinases
Cells
Cell Membrane
Dyneins
Kinesin
Fluorescence Microscopy
Recycling
Assays
Microscopic examination

Keywords

  • Dynein
  • Endosomes
  • Kinesin-1
  • Microtubules
  • ntcp
  • PKCζ
  • Recycling

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Structural Biology
  • Molecular Biology
  • Genetics

Cite this

PKCζ is required for microtubule-based motility of vesicles containing the ntcp transporter. / Sarkar, Souvik; Bananis, Eustratios; Nath, Sangeeta; Anwer, M. Sawkat; Wolkoff, Allan W.; Murray, John W.

In: Traffic, Vol. 7, No. 8, 08.2006, p. 1078-1091.

Research output: Contribution to journalArticle

Sarkar, Souvik ; Bananis, Eustratios ; Nath, Sangeeta ; Anwer, M. Sawkat ; Wolkoff, Allan W. ; Murray, John W. / PKCζ is required for microtubule-based motility of vesicles containing the ntcp transporter. In: Traffic. 2006 ; Vol. 7, No. 8. pp. 1078-1091.
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