Multistep regulation of autophagy by WNK1

Sachith Gallolu Kankanamalage, A. Young Lee, Chonlarat Wichaidit, Andres Lorente-Rodriguez, Akansha M. Shah, Steve Stippec, Angelique W. Whitehurst, Melanie H. Cobb

Research output: Contribution to journalArticle

10 Scopus citations

Abstract

The with-no-lysine (K) (WNK) kinases are an atypical family of protein kinases that regulate ion transport across cell membranes. Mutations that result in their overexpression cause hypertension-related disorders in humans. Of the four mammalian WNKs, only WNK1 is expressed throughout the body. We report that WNK1 inhibits autophagy, an intracellular degradation pathway implicated in several human diseases. Using small-interfering RNA-mediated WNK1 knockdown, we show autophagosome formation and autophagic flux are accelerated. In cells with reduced WNK1, basal and starvation-induced autophagy is increased. We also show that depletion of WNK1 stimulates focal class III phospha-tidylinositol 3-kinase complex (PI3KC3) activity, which is required to induce autophagy. Depletion of WNK1 increases the expression of the PI3KC3 upstream regulator unc-51-like kinase 1 (ULK1), its phos-phorylation, and activation of the kinase upstream of ULK1, the AMP-activated protein kinase. In addition, we show that the N-terminal region of WNK1 binds to the UV radiation resistance-associated gene (UVRAG) in vitro and WNK1 partially colocalizes with UVRAG, a component of a PI3KC3 complex. This colocalization decreases upon starvation of cells. Depletion of the SPS/STE20-related pro-line-alanine-rich kinase, a WNK1-activated enzyme, also induces autophagy in nutrient-replete or-starved conditions, but depletion of the related kinase and WNK1 substrate, oxidative stress responsive 1, does not. These results indicate that WNK1 inhibits autophagy by multiple mechanisms.

Original languageEnglish (US)
Pages (from-to)14342-14347
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume113
Issue number50
DOIs
Publication statusPublished - Dec 13 2016

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Keywords

  • PI3KC3
  • SPAK
  • ULK1
  • UVRAG
  • Vps34

ASJC Scopus subject areas

  • General

Cite this

Kankanamalage, S. G., Lee, A. Y., Wichaidit, C., Lorente-Rodriguez, A., Shah, A. M., Stippec, S., ... Cobb, M. H. (2016). Multistep regulation of autophagy by WNK1. Proceedings of the National Academy of Sciences of the United States of America, 113(50), 14342-14347. https://doi.org/10.1073/pnas.1617649113