Autosomal recessive bestrophinopathy is not associated with the loss of bestrophin-1 anion channel function in a patient with a novel BEST1 mutation

Adiv A. Johnson, Lori A. Bachman, Benjamin J. Gilles, Samuel D. Cross, Kimberly E. Stelzig, Zachary T. Resch, Lihua Y Marmorstein, Jose S Pulido, Alan D Marmorstein

Research output: Contribution to journalArticle

26 Scopus citations


PURPOSE. Mutations in BEST1, encoding bestrophin-1 (Best1), cause autosomal recessive bestrophinopathy (ARB). Encoding bestrophin-1 is a pentameric anion channel localized to the basolateral plasma membrane of the RPE. Here, we characterize the effects of the mutations R141H (CGC > CAC) and I366fsX18 (c.1098_1100+7del), identified in a patient in our practice, on Best1 trafficking, oligomerization, and channel activity. METHODS. Currents of Cl<sup>-</sup> were assessed in transfected HEK293 cells using whole-cell patch clamp. Best1 localization was assessed by confocal microscopy in differentiated, humaninduced pluripotent stem cell-derived RPE (iPSC-RPE) cells following expression of mutants via adenovirus-mediated gene transfer. Oligomerization was evaluated by coimmunoprecipitation in iPSC-RPE and MDCK cells. RESULTS. Compared to Best1, Best1<sup>I366fsX18</sup> currents were increased while Best1<sup>R141H</sup> Cl<sup>-</sup> currents were diminished. Coexpression of Best1<sup>R141H</sup> with Best1 or Best1I366fsX18 resulted in rescued channel activity. Overexpressed Best1, Best<sup>1R141H</sup>, and Best1<sup>I366fsX18</sup> were all properly localized in iPSC-RPE cells; Best1<sup>R141H</sup> and Best1<sup>I366fsX18</sup> coimmunoprecipitated with endogenous Best1 in iPSC-RPE cells and with each other in MDCK cells. CONCLUSIONS. The first 366 amino acids of Best1 are sufficient to mediate channel activity and homo-oligomerization. The combination of Best1 and Best1<sup>R141H</sup> does not cause disease, while Best1<sup>R141H</sup> together with Best1<sup>I366fsX18</sup> causes ARB. Since both combinations generate comparable Cl<sup>-</sup> currents, this indicates that ARB in this patient is not caused by a loss of channel activity. Moreover, Best1<sup>I366fsX18</sup> differs from Best1 in that it lacks most of the cytosolic C-terminal domain, suggesting that the loss of this region contributes significantly to the pathogenesis of ARB in this patient.

Original languageEnglish (US)
Pages (from-to)4619-4630
Number of pages12
JournalInvestigative Ophthalmology and Visual Science
Issue number8
StatePublished - 2015


  • Best1
  • iPSC-RPE
  • Localization
  • Oligomerization

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience

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