p21-activated kinases regulate actin remodeling in glomerular podocytes.
The tyrosine phosphorylation of nephrin is reported to regulate podocyte morphology via the Nck adaptor proteins. The Pak family of kinases are regulators of the actin cytoskeleton and are recruited to the plasma membrane via Nck. Here, we investigated the role of Pak in podocyte morphology. Pak1/2 were expressed in cultured podocytes. In mouse podocytes, Pak2 was predominantly phosphorylated, concentrated at the tips of the cellular processes, and its expression and/or phosphorylation were further increased when differentiated. Overexpression of rat nephrin in podocytes increased Pak1/2 phosphorylation, which was abolished when the Nck binding sites were mutated. Furthermore, dominant-negative Nck constructs blocked the Pak1 phosphorylation induced by antibody-mediated cross linking of nephrin. Transient transfection of constitutively kinase-active Pak1 into differentiated mouse podocytes decreased stress fibers, increased cortical F-actin, and extended the cellular processes, whereas kinase-dead mutant, kinase inhibitory construct, and Pak2 knockdown by shRNA had the opposite effect. In a rat model of puromycin aminonucleoside nephrosis, Pak1/2 phosphorylation was decreased in glomeruli, concomitantly with a decrease of nephrin tyrosine phosphorylation. These results suggest that Pak contributes to remodeling of the actin cytoskeleton in podocytes. Disturbed nephrin-Nck-Pak interaction may contribute to abnormal morphology of podocytes and proteinuria.
Zhu
Jianxin
J
Department of Medicine, McGill University Health Centre, 3775 University Street, Montreal, Quebec, Canada.
Attias
Ortal
O
Aoudjit
Lamine
L
Jiang
Ruihua
R
Kawachi
Hiroshi
H
Takano
Tomoko
T
D000199:Actins / Q000502:physiology*; D000818:Animals; D000903:Antibiotics, Antineoplastic / Q000009:adverse effects; D002460:Cell Line; D005786:Gene Expression Regulation / Q000502:physiology; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007527:Isoenzymes; D007678:Kidney Glomerulus / Q000166:cytology* / Q000378:metabolism; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D009401:Nephrosis / Q000139:chemically induced / Q000378:metabolism; D010766:Phosphorylation; D051179:Plakins / Q000502:physiology; D050199:Podocytes / Q000166:cytology* / Q000378:metabolism; D011692:Puromycin Aminonucleoside / Q000009:adverse effects; D051381:Rats; D054462:p21-Activated Kinases / Q000235:genetics / Q000378:metabolism*
D000199:Actins; D000903:Antibiotics, Antineoplastic; D007527:Isoenzymes; D008565:Membrane Proteins; D051179:Plakins; C113267:nephrin; D011692:Puromycin Aminonucleoside; D054462:p21-Activated Kinases