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Volume 272, Number 39, Issue of September 26, 1997 pp. 24530-24535
©1997 by The American Society for Biochemistry and Molecular Biology, Inc.

The AtVAM3 Encodes a Syntaxin-related Molecule Implicated in the Vacuolar Assembly in Arabidopsis thaliana

(Received for publication, June 2, 1997, and in revised form, July 28, 1997)

Masa H. Sato , Norihiro Nakamura , Yoshinori Ohsumi , Hiroshi Kouchi , Maki Kondo ^** , Ikuko Hara-Nishimura ^** , Mikio Nishimura ^** and Yoh Wada

From the  Department of Biology, Graduate School of Arts and Sciences, University of Tokyo, Komaba, Tokyo 153, the  Department of Applied Physiology, National Institute of Agrobiological Resources, Tsukuba 305, and the ^** Department of Cell Biology, National Institute for Basic Biology, Okazaki 444, Japan

The vacuole constitutes a large compartment in plant and fungal cells. The VAM3 gene of Saccharomyces cerevisiae encodes a syntaxin-related protein required for vacuolar assembly. An Arabidopsis thaliana cDNA library, designed for expression in S. cerevisiae, was screened for cDNAs able to complement defective vacuolar assembly of the vam3 mutation. One cDNA, encoding a 33-kDa protein with structural similarities to the other syntaxins, was identified. The product of AtVAM3 (AtVam3p) was expressed in various tissues including roots, leaves, inflorescence stems, flower buds, and young siliques. The AtVAM3 transcripts were abundant in undifferentiated cells in the meristematic region. AtVam3p fractionated predominantly to an 8,000 × g pellet fraction where a vacuolar membrane protein H⁺-translocating inorganic pyrophosphatase (H⁺-PPase) also fractionated. Immunoelectron microscopy showed that AtVam3p was localized to restricted regions on the vacuolar membranes. We propose that AtVam3p provides the t-SNARE function in the vacuolar assembly in A. thaliana.

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