Calcium Dependence of Polycystin-2 Channel Activity Is Modulated by Phosphorylation at Ser812

Yiqiang Cai, Georgia Anyatonwu, Dayne Okuhara, Kyu Beck Lee, Zhiheng Yu, Tamehito Onoe, Chang Lin Mei, Qi Qian, Lin Geng, Ralph Wiztgall, Barbara E. Ehrlich, Stefan Somlo

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Abstract

Polycystin-2 (PC-2) is a non-selective cation channel that, when mutated, results in autosomal dominant polycystic kidney disease. In an effort to understand the regulation of this channel, we investigated the role of protein phosphorylation in PC-2 function. We demonstrated the direct incorporation of phosphate into PC-2 in cells and tissues and found that this constitutive phosphorylation occurs at Ser812, a putative casein kinase II (CK2) substrate domain. Ser812 can be phosphorylated by CK2 in vitro and substitution S812A results in failure to incorporate phosphate in cultured epithelial cells. Non-phosphorylated forms of PC-2 traffic normally in the endoplasmic reticulum and cilial compartments and retain homo- and hetero-multimerization interactions with PC-2 and polycystin-1, respectively. Single-channel studies of PC-2, S812A, and a substitution mutant, T721A, not related to phosphorylation show that PC-2 and S812A function as divalent cation channels with similar current amplitudes across a range of holding potentials; the T721A channel is not functional. Channel open probabilities for PC-2 and S812A show a bell-shaped dependence on cytoplasmic Ca2+ but there is a shift in this Ca2+ dependence such that S812A is 10-fold less sensitive to Ca2+ activation/inactivation than the wild type PC-2 channel. In vivo analysis of PC-2-dependent enhanced intracellular Ca 2+ transients found that S812A resulted in enhanced transient duration and relative amplitude intermediate between control cells and those overexpressing wild type PC-2. Phosphorylation at Ser812 modulates PC-2 channel activity and factors regulating this phosphorylation are likely to play a role in the pathogenesis of polycystic kidney disease.

Original languageEnglish (US)
Pages (from-to)19987-19995
Number of pages9
JournalJournal of Biological Chemistry
Volume279
Issue number19
DOIs
StatePublished - May 7 2004

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Cai, Y., Anyatonwu, G., Okuhara, D., Lee, K. B., Yu, Z., Onoe, T., Mei, C. L., Qian, Q., Geng, L., Wiztgall, R., Ehrlich, B. E., & Somlo, S. (2004). Calcium Dependence of Polycystin-2 Channel Activity Is Modulated by Phosphorylation at Ser812. Journal of Biological Chemistry, 279(19), 19987-19995. https://doi.org/10.1074/jbc.M312031200