PIP5KIβ Selectively Modulates Apical Endocytosis in Polarized Renal Epithelial Cells

Szalinski, CM and Guerriero, CJ and Ruiz, WG and Docter, BE and Rbaibi, Y and Pastor-Soler, NM and Apodaca, G and Puthenveedu, MA and Weisz, OA (2013) PIP5KIβ Selectively Modulates Apical Endocytosis in Polarized Renal Epithelial Cells. PLoS ONE, 8 (1).

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Abstract

Localized synthesis of phosphatidylinositol 4,5-bisphosphate [PtdIns(4,5)P2] at clathrin coated pits (CCPs) is crucial for the recruitment of adaptors and other components of the internalization machinery, as well as for regulating actin dynamics during endocytosis. PtdIns(4,5)P2 is synthesized from phosphatidylinositol 4-phosphate by any of three phosphatidylinositol 5-kinase type I (PIP5KI) isoforms (α, β or γ). PIP5KIβ localizes almost exclusively to the apical surface in polarized mouse cortical collecting duct cells, whereas the other isoforms have a less polarized membrane distribution. We therefore investigated the role of PIP5KI isoforms in endocytosis at the apical and basolateral domains. Endocytosis at the apical surface is known to occur more slowly than at the basolateral surface. Apical endocytosis was selectively stimulated by overexpression of PIP5KIβ whereas the other isoforms had no effect on either apical or basolateral internalization. We found no difference in the affinity for PtdIns(4,5)P2-containing liposomes of the PtdIns(4,5)P2 binding domains of epsin and Dab2, consistent with a generic effect of elevated PtdIns(4,5)P2 on apical endocytosis. Additionally, using apical total internal reflection fluorescence imaging and electron microscopy we found that cells overexpressing PIP5KIβ have fewer apical CCPs but more internalized coated structures than control cells, consistent with enhanced maturation of apical CCPs. Together, our results suggest that synthesis of PtdIns(4,5)P2 mediated by PIP5KIβ is rate limiting for apical but not basolateral endocytosis in polarized kidney cells. PtdIns(4,5)P2 may be required to overcome specific structural constraints that limit the efficiency of apical endocytosis. © 2013 Szalinski et al.

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Creators	Email	Pitt Username	ORCID
Szalinski, CM
Guerriero, CJ	cjg11@pitt.edu	CJG11	0000-0002-1046-2300
Ruiz, WG	ruizw@pitt.edu	RUIZW
Docter, BE
Rbaibi, Y
Pastor-Soler, NM
Apodaca, G	gla6@pitt.edu	GLA6
Puthenveedu, MA
Weisz, OA	weisz@pitt.edu	WEISZ