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Plant and Cell Physiology Advance Access originally published online on October 29, 2007
Plant and Cell Physiology 2007 48(12):1737-1746; doi:10.1093/pcp/pcm146
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© The Author 2007. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org

HCF208, a Homolog of Chlamydomonas CCB2, is Required for Accumulation of Native Cytochrome b6 in Arabidopsis thaliana

Dagmar Lyska, Susanne Paradies, Karin Meierhoff* and Peter Westhoff

Institut für Entwicklungs- und Molekularbiologie der Pflanzen, Heinrich-Heine-Universität, Universitätsstr. 1, D-40225 Düsseldorf, Germany

*Corresponding author: E-mail, karin.meierhoff{at}uni-duesseldorf.de; Fax, +49-211-8114871.


  Abstract

The cytochrome b6 subunit of the cytochrome b6f complex is a multiheme protein. Two b-type hemes are bound non-covalently to the protein, whereas the third heme (heme cn) is covalently attached via an atypical thioether bond. To understand the maturation of cytochrome b6 and to identify the assisting factors, we characterized the ethyl methanesulfonate-induced nuclear mutant hcf208. This Arabidopsis mutant shows a high chlorophyll fluorescence phenotype and does not accumulate the major cytochrome b6f complex subunits. Transcript levels and patterns of the four major polypeptides of the complex are equal to those of the wild type. The mutant cytochrome b6 polypeptide shows a faster migration behavior in SDS–PAGE compared with the wild type and it has no peroxidase activity. The HCF208 locus was mapped and the gene was cloned. Sequence analysis revealed that HCF208 is a homolog of the Chlamydomonas reinhardtii CCB2 protein, which is a factor mediating attachment of heme cn to the cytochrome b6 polypeptide as part of a novel heme biogenesis pathway, called system IV. Blue Native PAGE revealed residual amounts of the cytochrome b6f complex dimer in hcf208; however, this form is unable to participate in electron transport reactions.

Keywords: Arabidopsis - Chloroplast - Cytochrome b6f complex - Heme attachment - Heme cn

Abbreviations: BAC, bacterial artificial chromosome; CCB: C, cofactor binding; C, cytochrome b6f complex; B, subunit PetB; EMS, ethyl methanesulfonate; HCF, high chlorophyll fluorescence; PFD, photon flux density

(Received August 10, 2007; Accepted October 23, 2007)
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