Molecular characterization of plasma membrane H+-ATPase in a carrot mutant cell line with enhanced citrate excretion

Takashi Ohno, Shinobu Nakahira, Yuji Suzuki, Tomomi Kani, Tetsuo Hara, Hiroyuki Koyama

Research output: Contribution to journalArticlepeer-review

17 Citations (Scopus)

Abstract

The citrate efflux of a carrot mutant cell line (IPG; insoluble phosphate grower) with enhanced citrate excretion is concomitant with an electrically equivalent H+ release. Furthermore, plasma membrane H +-ATPase activity (EC 3.6.1.35) in the IPG cell line is known to be greater than in the wild-type cell line. In the present study, molecular characterization of plasma membrane H+-ATPase in the IPG cell line was conducted to examine the involvement of this enzyme in enhanced citrate excretion. Four type-II (DcPA 1, 2, 4 and 5) and two type-I (DcPA 3 and 6) full-sized cDNAs encoding Daucus carota plasma membrane H+-ATPase were cloned. The transcript level of the most abundant isoform, DcPA 1, was three times higher in the IPG cell line than in the wild-type cells. The transcript levels of the other isoforms showed no differences between cell types. The plasma membrane H+-ATPase protein level in the IPG cell line was 1.6-fold that of the wild-type cell line when analysed by western blot analysis. These results indicate that the higher activity of plasma membrane H+-ATPase in the IPG cell line is due to up-regulation of DcPA 1. Antisense inhibition of the DcPA 1 gene also led to decreased citrate excretion from the transgenic IPG cell line, suggesting that the up-regulation of plasma membrane H+-ATPase is involved in enhanced citrate excretion.

Original languageEnglish
Pages (from-to)265-274
Number of pages10
JournalPhysiologia Plantarum
Volume122
Issue number2
DOIs
Publication statusPublished - 2004 Oct 1

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science
  • Cell Biology

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