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J. Biol. Chem., Vol. 279, Issue 7, 5429-5434, February 13, 2004

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Identification of Murr1 as a Regulator of the Human Epithelial Sodium Channel^*

Wolfgang Biasio, Tina Chang, C. Joy McIntosh, and Fiona J. McDonald

From the Department of Physiology, University of Otago, Dunedin 9001, New Zealand and the School of Biological Sciences, Victoria University, Wellington 6004, New Zealand

The human epithelial sodium channel (ENaC) subunit is related to the -, -, and ENaC subunits that control salt homeostasis. ENaC forms an amiloride-sensitive Na⁺ channel with the and subunits. However, the in vivo function of ENaC is not known. To gain insight into the function of ENaC, a yeast two-hybrid screen of a human brain cDNA library was carried out using the C- and N-terminal domains of ENaC. A novel ENaC-interacting protein called Murr1 (mouse U2af1-rs1 region) was isolated in the C-terminal domain screen. Murr1 is a 21-kDa protein mutated in Bedlington terriers suffering from copper toxicosis. The interaction of Murr1 and ENaC was confirmed by glutathione S-transferase pulldown assay and coimmunoprecipitation. To test the functional significance of the interaction, Murr1 was coexpressed with ENaC in Xenopus oocytes. Murr1 inhibited amiloride-sensitive sodium current in a dose-dependent manner. In addition, deletion of the last 59 amino acids of ENaC abolished the inhibition. Murr1 also bound to the - and ENaC subunits and inhibited ENaC sodium current. Therefore, these results suggest that Murr1 is a novel regulator of ENaC.

Received for publication, October 10, 2003 , and in revised form, November 11, 2003.

^* This work was supported by a grant from the Marsden Fund of New Zealand. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Physiology, University of Otago, Dunedin 9001, P. O. Box 913, New Zealand. Tel.: 64-3-4797329; Fax: 64-3-4797323; E-mail: fiona.mcdonald{at}stonebow.otago.ac.nz.

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