Abstract
BackgroundVacuolar type H+-ATPases play a critical role in the maintenance of vacuolar homeostasis in plant cells. V-ATPases are also involved in plants' defense against environmental stress. This research examined the expression and regulation of the catalytic subunit of the vacuolar type H+-ATPase in Arabidopsis thaliana and the effect of environmental stress on multiple transcripts generated by this gene.ResultsEvidence suggests that subunit A of the vacuolar type H+-ATPase is encoded by a single gene in Arabidopsis thaliana. Genome blot analysis showed no indication of a second subunit A gene being present. The single gene identified was shown by whole RNA blot analysis to be transcribed in all organs of the plant. Subunit A was shown by sequencing the 3' end of multiple cDNA clones to exhibit multi site polyadenylation. Four different poly (A) tail attachment sites were revealed. Experiments were performed to determine the response of transcript levels for subunit A to environmental stress. A PCR based strategy was devised to amplify the four different transcripts from the subunit A gene.ConclusionsAmplification of cDNA generated from seedlings exposed to cold, salt stress, and etiolation showed that transcript levels for subunit A of the vacuolar type H+-ATPase in Arabidopsis were responsive to stress conditions. Cold and salt stress resulted in a 2–4 fold increase in all four subunit A transcripts evaluated. Etiolation resulted in a slight increase in transcript levels. All four transcripts appeared to behave identically with respect to stress conditions tested with no significant differential regulation.
Highlights
Vacuolar type H+-ATPases play a critical role in the maintenance of vacuolar homeostasis in plant cells
The primary nucleotide and amino acid sequence of the A. thaliana subunit A cDNA showed a high degree of identity with subunit A cDNA sequenced from other organisms
Genome blot analysis was utilized to determine the number of genes in the A. thaliana genome corresponding to subunit A of the vacuolar type ATPase
Summary
Vacuolar type H+-ATPases play a critical role in the maintenance of vacuolar homeostasis in plant cells. Vacuolar-type H+-ATPases are enzymes responsible for the energization of membranes and the acidification of compartments within the eukaryotic cell via the establishment of proton and electrochemical gradients at the expense of ATP. The vacuolar type H+-ATPase in plants is a large multimeric enzyme complex whose function is to pump protons across a membrane via primary active transport. Vacuolar type H+-ATPases are critical for the maintenance of homeostasis in eukaryotic cells [10,11]. The vacuolar type ATPase is thought to be primarily responsible for the acidification and expansion of the large central vacuole [12,13,14,15]. The influx of water and metabolites into the vacuole is dependent in part on the generation of a proton motive force across the tonoplast
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
