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Journal of Experimental Botany, Vol. 53, No. 378, pp. 2273-2275, November 1, 2002
© 2002 Oxford University Press

Functional expression of Acetabularia acetabulum vacuolar H+-pyrophosphatase in a yeast VMA3-deficient strain

Received 31 May 2002; Accepted 4 July 2002

Mikiko Ikeda4,2, Kimiko Umami2, Misato Hinohara2, Yoshie Tanimura2, Atsuko Ohmae2, Yoichi Nakanishi3 and Masayoshi Maeshima3

2 Department of Nutritional Science, Faculty of Health and Welfare Science, Okayama Prefectural University, Soja 719-1197, Japan
3 Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan

Abbreviations: V-PPase, vacuolar H+-pyrophosphatase; V-ATPase, vacuolar H+-ATPase; VMA3, gene coding for the proteolipid subunit of Saccharomyces cerevisiae V-ATPase.

The function of the translation product of cDNA for Acetabularia vacuolar H+-pyrophosphatase was examined using the Saccharomyces cerevisiae VMA3-deficient strain. The open reading frame of Acetabularia H+-pyrophosphatase was revealed to encode 751 amino acids (721 or 751 amino acids in a previous paper). The acidification of the vacuole was observed by fluorescence microscopy when the cDNA was constructed in pYES2. Immunoblot analysis also supported the localization of the translation product in the vacuolar-membrane-enriched fraction.

Key words: Key words: Acetabularia acetabulum, heterologous expression, proton transport, vacuolar H+-pyrophosphatase, yeast.


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