Characterization of the V-type H((+))-ATPase in the resurrection plant Tortula ruralis: accumulation and polysomal recruitment of the proteolipid c subunit in response to salt-stress.

Tortula ruralis is an important experimental system for the study of plant vegetative desiccation tolerance. EST gene discovery efforts utilizing desiccated gametophytes have identified a cDNA Vac1 encoding a predicted polypeptide with significant similarity to the vacuolar H(+)-ATPase c subunit. VAC1, the 167 amino acid deduced polypeptide, has a predicted molecular mass of 16.9 kDa, and a predicted pI of 9.7. Phylogenetic analysis demonstrated that previously characterized proteolipid polypeptide sequences could be reproducibly grouped into two major clades and that VAC1 forms a discrete evolutionary group. RNA blot and Western blot hybridizations were used to analyse expression of Vac1 and accumulation of VAC1 in response to (1) desiccation and rehydration, (2) increased NaCl concentration, and (3) NaCl-shock. During a desiccation-rehydration cycle, Vac1 transcripts are expressed in both the total and polysomal RNA fractions in approximately equal amounts, and the steady-state transcript levels are unchanged. However, Vac1 transcript levels increased in response to both elevated NaCl concentration and NaCl-shock. There is a preferential accumulation of Vac1 transcripts within the polysomal RNA fraction in response to salt stress, and these data suggest that T. ruralis possesses a salinity-stress-dependent and desiccation-stress-independent mechanism for post-transcriptional gene control. Using a cotton anti-c subunit polyclonal antibody raised against the C-terminal domain, it was shown that the amount of Tortula 16 kDa proteolipid in the tonoplast protein fraction was unaffected by any stress treatment.