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JXB Advance Access originally published online on July 7, 2007
Journal of Experimental Botany 2007 58(11):2839-2849; doi:10.1093/jxb/erm094
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© The Author [2007]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

RESEARCH PAPER

Cloning of the PpNHAD1 transporter of Physcomitrella patens, a chloroplast transporter highly conserved in photosynthetic eukaryotic organisms

Javier Barrero-Gil, Alonso Rodríguez-Navarro and Begoña Benito*

Departamento de Biotecnología, Universidad Politécnica de Madrid, 28040 Madrid, Spain

* To whom correspondence should be addressed. E-mail: begona.benito{at}upm.es

A cDNA that encodes a transporter from the NHAD family was identified in Physcomitrella patens. Computer-based searches using the amino acid sequence of PpNHAD1 revealed that, in addition to being expressed in flowering plants, highly conserved transporters of this family are expressed in red algae, green algae, mosses, liverworts, and photosynthetic stramenopiles, but not in heterotrophic stramenopiles. A chloroplast transit peptide was detected in PpNHAD1 and in most of the related sequences, indicating that PpNHAD1 is a chloroplast transporter. A PpNHAD1-GFP fusion localized to the chloroplast in Physcomitrella protoplasts, and truncation of the N-terminus of the protein dispersed the fluorescence signal outside the chloroplast. PpNHAD1 did not show functional expression in either yeast or bacterial mutants, but truncated proteins with shorter N-termini, PpNHAD1-1 and PpNHAD1-2, could be functionally expressed in bacteria. PpNHAD1-1 alleviated the Li+ intolerance of a Na+-efflux Escherichia coli mutant at acidic pH values. Both PpNHAD1-1 and PpNHAD1-2 reduced the K+ requirements of a K+-influx E. coli mutant more actively at high pH values. PpNHAD1 seems to be an important transporter that mediates ionic homeostasis in chloroplasts from red algae to flowering plants.

Key words: Antiporter, chloroplast, lithium, NHAD, Physcomitrella, potassium, sodium

Received 25 October 2006; Revised 26 March 2007 Accepted 4 April 2007


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