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First published online May 4, 2004
doi: 10.1242/10.1242/jcs.01092

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Chloroplast division site placement requires dimerization of the ARC11/AtMinD1 protein in Arabidopsis

Makoto T. Fujiwara^1,2,, Ayako Nakamura², Ryuuichi Itoh^2,*, Yukihisa Shimada², Shigeo Yoshida² and Simon Geir Møller^1,

¹ Department of Biology, University of Leicester, University Road, Leicester, LE1 7RH, UK
² Plant Functions Laboratory and Plant Science Center, RIKEN, Hirosawa 2-1, Wako, Saitama 351-0198, Japan

Authors for correspondence (e-mail: mtf1{at}mac.com; sgm5{at}le.ac.uk)

Accepted 9 January 2004

Chloroplast division is mediated by the coordinated action of a prokaryote-derived division system(s) and a host eukaryote-derived membrane fission system(s). The evolutionary conserved prokaryote-derived system comprises several nucleus-encoded proteins, two of which are thought to control division site placement at the midpoint of the organelle: a stromal ATPase MinD and a topological specificity factor MinE. Here, we show that arc11, one of 12 recessive accumulation and replication of chloroplasts (arc) mutants in Arabidopsis, contains highly elongated and multiple-arrayed chloroplasts in developing green tissues. Genomic sequence analysis revealed that arc11 contains a missense mutation in -helix 11 of the chloroplast-targeted AtMinD1 changing an Ala at position 296 to Gly (A296G). Introduction of wild-type AtMinD1 restores the chloroplast division defects of arc11 and quantitative RT-PCR analysis showed that the degree of complementation was highly dependent on transgene expression levels. Overexpression of the mutant ARC11/AtMinD1 in transgenic plants results in the inhibition of chloroplast division, showing that the mutant protein has retained its division inhibition activity. However, in contrast to the defined and punctate intraplastidic localization patterns of an AtMinD1-YFP fusion protein, the single A296G point mutation in ARC11/AtMinD1 results in aberrant localization patterns inside chloroplasts. We further show that AtMinD1 is capable of forming homodimers and that this dimerization capacity is abolished by the A296G mutation in ARC11/AtMinD1. Our data show that arc11 is a loss-of-function mutant of AtMinD1 and suggest that the formation of functional AtMinD1 homodimers is paramount for appropriate AtMinD1 localization, ultimately ensuring correct division machinery placement and chloroplast division in plants.

Key words: Arabidopsis, Chloroplast division, Min system, arc mutant

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