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A functional study of plasma-membrane calcium-pump isoform 2 mutants causing digenic deafness
- Ficarella, R., Di Leva, F., Bortolozzi, M., Ortolano, S., Donaudy, F., Petrillo, M., Melchionda, S., Lelli, A., Domi, T., Fedrizzi, L., Lim, D., Shull, G.E., Gasparini, P., Brini, M., Mammano, F., Carafoli, E.
- Proceedings of the National Academy of Sciences of the United States of America 2007 v.104 no.5 pp. 1516-1521
- cadherins, calcium, deafness, genes, humans, image analysis, knockout mutants, mice, missense mutation, plasma membrane, variance
- Ca²⁺ enters the stereocilia of hair cells through mechanoelectrical transduction channels opened by the deflection of the hair bundle and is exported back to endolymph by an unusual splicing isoform (w/a) of plasma-membrane calcium-pump isoform 2 (PMCA2). Ablation or missense mutations of the pump cause deafness, as described for the G283S mutation in the deafwaddler (dfw) mouse. A deafness-inducing missense mutation of PMCA2 (G293S) has been identified in a human family. The family also was screened for mutations in cadherin 23, which accentuated hearing loss in a previously described human family with a PMCA2 mutation. A T1999S substitution was detected in the cadherin 23 gene of the healthy father and affected son but not in that of the unaffected mother, who presented instead the PMCA2 mutation. The w/a isoform was overexpressed in CHO cells. At variance with the other PMCA2 isoforms, it became activated only marginally when exposed to a Ca²⁺ pulse. The G293S and G283S mutations delayed the dissipation of Ca²⁺ transients induced in CHO cells by InsP₃. In organotypic cultures, Ca²⁺ imaging of vestibular hair cells showed that the dissipation of stereociliary Ca²⁺ transients induced by Ca²⁺ uncaging was compromised in the dfw and PMCA2 knockout mice, as was the sensitivity of the mechanoelectrical transduction channels to hair bundle displacement in cochlear hair cells.