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Potential role of the rice OsCCS52A gene in endoreduplication

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Abstract

In eukaryotes, the cell cycle consists of four distinct phases: G1, S, G2 and M. In certain condition, the cells skip M-phase and undergo endoreduplication. Endoreduplication, occurring during a modified cell cycle, duplicates the entire genome without being followed by M-phase. A cycle of endoreduplication is common in most of the differentiated cells of plant vegetative tissues and it occurs extensively in cereal endosperm cells. Endoreduplication occurs when CDK/Cyclin complex low or inactive caused by ubiquitin-mediated degradation by APC and their activators. In this study, rice cell cycle switch 52 A (OsCCS52A), an APC activator, is functionally characterized using the reverse genetic approach. In rice, OsCCS52A is highly expressed in seedlings, flowers, immature panicles and 15 DAP kernels. Localization studies revealed that OsCCS52A is a nuclear protein. OsCCS52A interacts with OsCdc16 in yeast. In addition, overexpression of OsCCS52A inhibits mitotic cell division and induces endoreduplication and cell elongation in fission yeast. The homozygous mutant exhibits dwarfism and smaller seeds. Further analysis demonstrated that endoreduplication cycles in the endosperm of mutant seeds were disturbed, evidenced by reduced nuclear and cell sizes. Taken together, these results suggest that OsCCS52A is involved in maintaining normal seed size formation by mediating the exit from mitotic cell division to enter the endoreduplication cycles in rice endosperm.

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Abbreviations

APC:

Anaphase-promoting complex

CDK:

Cyclin-dependent kinase

DAP:

Days after pollination

DAPI:

4′,6-Diamidino-2-phenylindole

EGFP:

Enhanced-gene fluorescent protein

OsCCS52A :

Rice cell cycle switch 52 A

OsCdc16:

Rice cell division cycle protein 16

M-phase:

Mitotic phase

MS:

Murashige and Skoog

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea funded by the Ministry of Education, Science and Technology (2009-0068189). MS, JYC, MHJ and NE were supported by scholarships from the BK21 program, MEST, Korea.

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Correspondence to Young-Min Woo or Daeyoung Son.

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Supplementary Tab. S1 List of oligonucleotides used in this study.

Supplementary Fig. S1 Multiple sequence alignment and phylogenetic tree of closely-related CCS52 genes.

Amino acid sequences of OsCCS52A and the closest CCS52 members of rice (OsCCS52A and OsCCS52B), Medicago (MtCCS52A and MtCCS52B), Arabidopsis (AtCCS52A1, AtCCS52A2 and AtCCS52B), maize (ZmCCS52A and ZmCCS52B), human (HsCdh1) and fission yeast (SpSrw1) were aligned using ClustalX program. Numbers at the right indicate the amino acid residue position. The conserved APC-interacting motifs (C-box, CSM, and the C-terminal IR residues) are boxed in the N-terminus and the C-terminus, respectively; and the seven WD40-repeats are indicated by a double-headed arrow. Gaps (-) were introduced for maximum alignment.

Supplementary Fig. S2 Phylogenetic tree of OsCCS52A (red underlined) and its closest members was constructed using Clustal X and MEGA 4 software.

Supplementary Fig. S3 Microscopy of the cross sections of mature leaves. Paraffin-embedded mature leaves of wild-type (left) and osccs52a-1 (right) were sectioned and stained with DAPI. Numbers indicate close-up images at the same portions of wild-type and osccs52a-1.

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Su’udi, M., Cha, JY., Jung, M.H. et al. Potential role of the rice OsCCS52A gene in endoreduplication. Planta 235, 387–397 (2012). https://doi.org/10.1007/s00425-011-1515-8

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