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Functional dependence on calcineurin by variants of the Saccharomyces cerevisiae vacuolar Ca 2+ /H + exchanger Vcx1p
Author(s) -
Pittman Jon K.,
Cheng NingHui,
Shigaki Toshiro,
Kunta Madhurababu,
Hirschi Kendal D.
Publication year - 2004
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.2004.04332.x
Subject(s) - calcineurin , saccharomyces cerevisiae , biology , vacuole , cytosol , biochemistry , phosphatase , mutant , ion transporter , yeast , membrane transport , sodium–hydrogen antiporter , microbiology and biotechnology , biophysics , membrane , chemistry , phosphorylation , sodium , enzyme , gene , cytoplasm , medicine , surgery , organic chemistry , transplantation
Summary The Ca 2+ ‐dependent protein phosphatase calcineurin is an important regulator of ion transporters from many organisms, including the Saccharomyces cerevisiae vacuolar Ca 2+ /H + exchanger Vcx1p. In yeast and plants, cation/H + exchangers are important in shaping cytosolic Ca 2+ levels involved in signal transduction and providing tolerance to potentially toxic concentrations of cations such as Ca 2+ , Mn 2+ and Cd 2+ . Previous genetic evidence suggested Vcx1p is negatively regulated by calcineurin. By utilizing direct transport measurements into vacuolar membrane vesicles, we demonstrate that Vcx1p is a low‐affinity Ca 2+ transporter and may also function in Cd 2+ transport, but cannot transport Mn 2+ . Furthermore, direct Ca 2+ transport by Vcx1p is calcineurin sensitive. Using a yeast growth assay, a mutant allele of VCX1 ( VCX1 ‐S204A/L208P), termed VCX1‐M1 , was previously found to confer strong Mn 2+ tolerance. Here we demonstrate that this Mn 2+ tolerance is independent of the Ca 2+ /Mn 2+ ‐ATPase Pmr1p and results from Mn 2+ ‐specific vacuolar transport activity of Vcx1‐M1p. This Mn 2+ transport by Vcx1‐M1p is calcineurin dependent, although the localization of Vcx1‐M1p to the vacuole appears to be calcineurin independent. Additionally, we demonstrate that mutation of L208P alone is enough to confer calcineurin‐dependent Mn 2+ tolerance. This study demonstrates that calcineurin can positively regulate the transport of cations by VCX1‐M1p.