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Plant J. 2019 Oct 22. doi: 10.1111/tpj.14579. [Epub ahead of print]

The formation of perinucleolar bodies is important for normal leaf development and requires the zinc-finger DNA-binding motif in Arabidopsis ASYMMETRIC LEAVES2.

Author information

1
Division of Biological Science, Graduate School of Science, Nagoya University, Nagoya, Aichi, 464-8602, Japan.
2
Graduate School of Bioscience and Biotechnology, Chubu University, Kasugai, Aichi, 487-8501, Japan.
3
Department of Applied Biological Science, Faculty of Science and Technology, Tokyo University of Science, Noda, Chiba, 278-8510, Japan.
4
Department of Biological Sciences, Graduate School of Science, Osaka University, 1-1 Machikaneyama-cho, Toyonaka, Osaka, 560-0043, Japan.
5
Central Research Institute, Ishihara Sangyo Kaisha, Ltd., 2-3-1 Nishi-Shibukawa, Kusatsu, Shiga, 525-0025, Japan.
6
Graduate School of Medical Sciences, Kanazawa University, Kakuma-machi, Kanazawa, Ishikawa, 920-1192, Japan.
7
JST, PRESTO, Furo-cho, Chikusa-ku, Nagoya, Aichi, 464-8601, Japan.
8
Institute of Transformative Bio-Molecules (ITbM), Nagoya University, Furo-cho, Chiku00sa-ku, Nagoya, Aichi, 464-8601, Japan.
9
Department of Biological Sciences, Graduate School of Science, University of Tokyo, 7-3-1 Hongo, Bukyo-ku, Tokyo, 113-0033, Japan.
10
Division of Biological Sciences, Faculty of Science, Hokkaido University, Sapporo, 060-0810, Japan.
11
Department of Biology, Faculty of Agriculture and Life Science, Hirosaki University, 3 Bunkyo-cho, Hirosaki, 036-8561, Japan.

Abstract

In Arabidopsis, the ASYMMETRIC LEAVES2 (AS2) protein plays a key role in the formation of flat symmetric leaves via direct repression of the abaxial gene ETT/ARF3. AS2 encodes a plant-specific nuclear protein that contains the AS2/LOB domain, which includes a zinc-finger (ZF) motif that is conserved in the AS2/LOB family. We have shown that AS2 binds to the coding DNA of ETT/ARF3, which requires the ZF motif. AS2 is co-localized with AS1 in perinucleolar bodies (AS2 bodies). To identify the amino acid signals in AS2 required for formation of AS2 bodies and function(s) in leaf formation, we constructed recombinant DNAs that encoded mutant AS2 proteins fused to yellow fluorescent protein. We examined the subcellular localization of these proteins in cells of cotyledons and leaf primordia of transgenic plants and cultured cells. The amino acid signals essential for formation of AS2 bodies were located within and adjacent to the ZF motif. Mutant AS2 that failed to form AS2 bodies also failed to rescue the as2-1 mutation. Our results suggest the importance of the formation of AS2 bodies and the nature of interactions of AS2 with its target DNA and nucleolar factors including NUCLEOLIN1. The partial overlap of AS2 bodies with perinucleolar chromocenters with condensed ribosomal RNA genes implies a correlation between AS2 bodies and the chromatin state. Patterns of AS2 bodies in cells during interphase and mitosis in leaf primordia were distinct from those in cultured cells, suggesting that the formation and distribution of AS2 bodies are developmentally modulated in plants.

KEYWORDS:

45S ribosomal RNA genes; ETTIN/AUXIN RESEPONSE FACTOR3; chromocenter; epigenetic factor AS2; nucleolus; perinucleolar body; zinc-finger motif

PMID:
31639235
DOI:
10.1111/tpj.14579

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