Cloning of the V‐ATPase subunit G in plant: functional expression and sub‐cellular localization

FEBS Letters

Rouquié, David¹  Tournaire-Roux, Colette¹  Szponarski, Wojciech¹  Doumas, Patrick¹  Rossignol, Michel¹ 

[1] Biochimie et Physiologie Moléculaire des Plantes, INRA/ENSA-M/CNRS URA 2133, Place Viala, 34060 Montpellier Cedex 1, France

关键词: H+-Translocating vacuolar-type ATPase;    Subcellular localization;    cDNA cloning;    Yeast complementation;    Nicotiana tabacum;   

DOI  :  10.1016/S0014-5793(98)01252-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

A 13-kDa tobacco plasma membrane protein was isolated from two-dimensional electrophoresis gels. After microsequencing, RT-PCR techniques and cDNA library screening allowed for the cloning of two cDNAs. These cDNAs encoded for the subunit G of the vacuolar H⁺-ATPase, the first one identified in plants. Analysis of mRNA distribution showed a maximum level in the leaves and in the stem of the apical part of the tobacco plant. Heterologous functional complementation of the yeast mutant (Δvma10::URA3) was achieved with the two cDNAs. After fractionation of microsomal membranes on linear sucrose gradient, Western blots were performed using antibodies against recombinant protein and three peaks were identified: one which comigrated with the tonoplast marker and the others at slightly higher density corresponding to endoplasmic reticulum and to plasma membrane fractions.

【 授权许可】

Unknown