Motor protein Myo1c is a podocyte protein that facilitates the transport of slit diaphragm protein Neph1 to the podocyte membrane

E. Arif, Mark Wagner, D. B. Johnstone, H. N. Wong, B. George, P. A. Pruthi, M. J. Lazzara, D. Nihalani

Research output: Contribution to journalArticle

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Abstract

The podocyte proteins Neph1 and nephrin organize a signaling complex at the podocyte cell membrane that forms the structural framework for a functional glomerular filtration barrier. Mechanisms regulating the movement of these proteins to and from the membrane are currently unknown. This study identifies a novel interaction between Neph1 and the motor protein Myo1c, where Myo1c plays an active role in targeting Neph1 to the podocyte cell membrane. Using in vivo and in vitro experiments, we provide data supporting a direct interaction between Neph1 and Myo1c which is dynamic and actin dependent. Unlike wild-type Myo1c, the membrane localization of Neph1 was significantly reduced in podocytes expressing dominant negative Myo1c. In addition, Neph1 failed to localize at the podocyte cell membrane and cell junctions in Myo1c-depleted podocytes. We further demonstrate that similarly to Neph1, Myo1c also binds nephrin and reduces its localization at the podocyte cell membrane. A functional analysis of Myo1c knockdown cells showed defects in cell migration, as determined by a wound assay. In addition, the ability to form tight junctions was impaired in Myo1c knockdown cells, as determined by transepithelial electric resistance (TER) and bovine serum albumin (BSA) permeability assays. These results identify a novel Myo1c-dependent molecular mechanism that mediates the dynamic organization of Neph1 and nephrin at the slit diaphragm and is critical for podocyte function.

Original languageEnglish
Pages (from-to)2134-2150
Number of pages17
JournalMolecular and Cellular Biology
Volume31
Issue number10
DOIs
StatePublished - May 2011

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Podocytes
Protein Transport
Diaphragm
Membranes
Proteins
Cell Membrane
Glomerular Filtration Barrier
Intercellular Junctions
Tight Junctions
Bovine Serum Albumin
Electric Impedance
Cell Movement
Actins
Permeability
Membrane Proteins
Wounds and Injuries

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Motor protein Myo1c is a podocyte protein that facilitates the transport of slit diaphragm protein Neph1 to the podocyte membrane. / Arif, E.; Wagner, Mark; Johnstone, D. B.; Wong, H. N.; George, B.; Pruthi, P. A.; Lazzara, M. J.; Nihalani, D.

In: Molecular and Cellular Biology, Vol. 31, No. 10, 05.2011, p. 2134-2150.

Research output: Contribution to journalArticle

Arif, E. ; Wagner, Mark ; Johnstone, D. B. ; Wong, H. N. ; George, B. ; Pruthi, P. A. ; Lazzara, M. J. ; Nihalani, D. / Motor protein Myo1c is a podocyte protein that facilitates the transport of slit diaphragm protein Neph1 to the podocyte membrane. In: Molecular and Cellular Biology. 2011 ; Vol. 31, No. 10. pp. 2134-2150.
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