ZMVHA-B1, the gene for subunit B of vacuolar H+-ATPase from the eelgrass Zostera marina L. Is able to replace vma2 in a yeast null mutant.
A vacuolar H(+)-ATPase (VHA) gene (ZMVHA-B1) was isolated from an eelgrass (Zostera marina) leaf cDNA library and was characterized to be approximately 1.4 kbp in length and to encode the B subunit protein of VHA comprising 488 amino acids. ZMVHA-B1 was highly expressed in all organs of eelgrass; the expression level was highest in the leaves. On transformation of a yeast vma2 null mutant with ZMVHA-B1, yeast cells became able to grow at pH 7.5, accompanied by the vesicular accumulation of LysoSensor green DND-189. Thus, ZMVHA-B1 expressed in yeast cells produced a functional B subunit that was efficiently incorporated into the VHA complex and eventually restored vacuolar morphology and activity. This success expedites the application of heterologous expression in yeast mutant cells to the screening of eelgrass genes involved in salt-resistance mechanisms, which are to be utilized in improving important crops.
Alemzadeh
Abbas
A
Department of Molecular Biotechnology, Graduate School of Advanced Sciences of Matter, Hiroshima University, 1-3-1 Kagamiyama, Higashi-Hiroshima 739-8530, Japan.
Fujie
Makoto
M
Usami
Shoji
S
Yoshizaki
Tsukasa
T
Oyama
Keiji
K
Kawabata
Toyoki
T
Yamada
Takashi
T
D003001:Cloning, Molecular; D004463:Ecology; D005816:Genetic Complementation Test; D007527:Isoenzymes / Q000235:genetics; D009154:Mutation; D021122:Protein Subunits; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012965:Sodium Chloride / Q000494:pharmacology; D025262:Vacuolar Proton-Translocating ATPases / Q000235:genetics* / Q000502:physiology; D035962:Zosteraceae / Q000201:enzymology*
D007527:Isoenzymes; D021122:Protein Subunits; D012965:Sodium Chloride; D025262:Vacuolar Proton-Translocating ATPases