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SYSTEMS BIOLOGY, MOLECULAR BIOLOGY, AND GENE REGULATION

Redundant Requirement for a Pair of PROTEIN ARGININE METHYLTRANSFERASE4 Homologs for the Proper Regulation of Arabidopsis Flowering Time^1,[C],[OA]

State Key Laboratory of Plant Genomics and National Center for Plant Gene Research, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (L.N., Y.Z., Y.P., C.L., X.C.); Graduate School of the Chinese Academy of Sciences, Beijing 100039, China (L.N., Y.Z.); and College of Life Science and Technology, Shanxi University, Taiyuan 030006, China (Y.P.)

CARM1/PRMT4 (for COACTIVATOR-ASSOCIATED ARGININE METHYLTRANSFERASE1/PROTEIN ARGININE METHYLTRANSFERASE4) catalyzes asymmetric dimethylation on arginine (Arg), and its functions in gene regulation is understood only in animal systems. Here, we describe AtPRMT4a and AtPRMT4b as a pair of Arabidopsis (Arabidopsis thaliana) homologs of mammalian CARM1/PRMT4. Recombinant AtPRMT4a and AtPRMT4b could asymmetrically dimethylate histone H3 at Arg-2, Arg-17, Arg-26, and myelin basic protein in vitro. Both AtPRMT4a and AtPRMT4b exhibited nuclear as well as cytoplasmic distribution and were expressed ubiquitously in all tissues throughout development. Glutathione S-transferase pull-down assays revealed that AtPRMT4a and AtPRMT4b could form homodimers and heterodimers in vitro, and formation of the heterodimer was further confirmed by bimolecular fluorescence complementation. Simultaneous lesions in AtPRMT4a and AtPRMT4b genes led to delayed flowering, whereas single mutations in either AtPRMT4a or AtPRMT4b did not cause major developmental defects, indicating the redundancy of AtPRMT4a and AtPRMT4b. Genetic analysis also indicated that atprmt4a atprmt4b double mutants phenocopied autonomous pathway mutants. Finally, we found that asymmetric methylation at Arg-17 of histone H3 was greatly reduced in atprmt4a atprmt4b double mutants. Taken together, our results demonstrate that AtPRMT4a and AtPRMT4b are required for proper regulation of flowering time mainly through the FLOWERING LOCUS C-dependent pathway.

² These authors contributed equally to the article.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Xiaofeng Cao (xfcao{at}genetics.ac.cn).

^[C] Some figures in this article are displayed in color online but in black and white in the print edition.

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www.plantphysiol.org/cgi/doi/10.1104/pp.108.124727

^* Corresponding author; e-mail xfcao{at}genetics.ac.cn.

Received June 13, 2008; accepted July 17, 2008; published July 25, 2008.

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