Promotion of neurite extension by protrudin requires its interaction with vesicle-associated membrane protein-associated protein

Shotaro Saita, Michiko Shirane, Tohru Natume, Shun Ichiro Iemura, Keiichi I. Nakayama

Research output: Contribution to journalArticle

43 Citations (Scopus)

Abstract

Protrudin is a protein that contains a Rab11-binding domain and a FYVE (lipid-binding) domain and that functions to promote neurite formation through interaction with the GDP-bound form of Rab11. Protrudin also contains a short sequence motif designated FFAT (two phenylalanines in an acidic tract), which in other proteins has been shown to mediate binding to vesicle-associated membrane protein-associated protein (VAP). We now show that protrudin associates and colocalizes with VAP-A, an isoform of VAP expressed in the endoplasmic reticulum. Both the interaction between protrudin and VAP-A as well as the induction of process formation by protrudin were markedly inhibited by mutation of the FFAT motif. Furthermore, depletion of VAP-A by RNA interference resulted in mislocalization of protrudin as well as in inhibition of neurite outgrowth induced by nerve growth factor in rat pheochromocytoma PC12 cells. These defects resulting from depletion of endogenous rat VAP-A in PC12 cells were corrected by forced expression of (RNA interference-resistant) human VAP-A but not by VAP-A mutants that have lost the ability to interact with protrudin. These results suggest that VAP-A is an important regulator both of the subcellular localization of protrudin and of its ability to stimulate neurite outgrowth.

Original languageEnglish
Pages (from-to)13766-13777
Number of pages12
JournalJournal of Biological Chemistry
Volume284
Issue number20
DOIs
Publication statusPublished - May 15 2009

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R-SNARE Proteins
Neurites
PC12 Cells
RNA Interference
Rats
RNA
Proteins
Pheochromocytoma
Nerve Growth Factor
Phenylalanine
Endoplasmic Reticulum
Protein Isoforms
Lipids
Defects
Mutation
Neuronal Outgrowth

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Promotion of neurite extension by protrudin requires its interaction with vesicle-associated membrane protein-associated protein. / Saita, Shotaro; Shirane, Michiko; Natume, Tohru; Iemura, Shun Ichiro; Nakayama, Keiichi I.

In: Journal of Biological Chemistry, Vol. 284, No. 20, 15.05.2009, p. 13766-13777.

Research output: Contribution to journalArticle

Saita, Shotaro ; Shirane, Michiko ; Natume, Tohru ; Iemura, Shun Ichiro ; Nakayama, Keiichi I. / Promotion of neurite extension by protrudin requires its interaction with vesicle-associated membrane protein-associated protein. In: Journal of Biological Chemistry. 2009 ; Vol. 284, No. 20. pp. 13766-13777.
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