Plant Physiol. Journal of Pharmacology and Experimental Therapeutics
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Published on February 25, 2009; 10.1104/pp.109.135582

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Received January 12, 2009
Accepted February 18, 2009

Preferential up-regulation of G2/M phase-specific genes by overexpression of the hyperactive form of NtmybA2 lacking its negative regulation domain in tobacco BY2 cells

Kiichi Kato , Ivan Galis , Shiori Suzuki , Satoshi Araki , Taku Demura , Marie-Claire Criqui , Thomas Potuschak , Pascal Genschik , Hiroo Fukuda , Ken Matsuoka , and Masaki Ito *

Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa, Nagoya, 464-8601 Japan; RIKEN Plant Science Center, Yokohama, Kanagawa, 230-0045 Japan; Central Research Institute, Ishihara Sangyo Kaisha, Ltd, 2-3-1 Nishi-shibukawa, Kusatsu, Shiga, 525-0025 Japan; Institut de Biologie Moleculaire des Plantes, Laboratoire Propre du Centre National de la Recherche Scientifique, Unite Propre de Recherche 2357, Conventionne avec l'Universite Louis Pasteur, 67084 Strasbourg, France

* Corresponding author; email: masakito{at}agr.nagoya-u.ac.jp.

Many G2/M phase-specific genes in plants contain mitosis-specific activator (MSA) elements, which act as G2/M phase-specific enhancers and bind with R1R2R3-Myb transcription factors. Here, we examined the genome-wide effects of NtmybA2 overexpression, one of the R1R2R3-Myb transcription factors in tobacco (Nicotiana tabacum). We used a custom-made 16K cDNA microarray for comparative transcriptome analysis of transgenic tobacco BY2 cell lines that overexpress NtmybA2 or its truncated hyperactive form. The microarray was also used to determine the transcript profile during the cell cycle in synchronized cultures of BY2 cells. Combined microarray data from transgenic lines and synchronized cells revealed that overexpression of the truncated hyperactive form of NtmybA2, but not its full-length form, preferentially up-regulated many G2/M phase-specific genes in BY2 cells. We determined promoter sequences of several such up-regulated genes and showed that all contain MSA-like motifs in the proximal regions of their promoters. One of the up-regulated genes, NtE2C, encoding for cyclin-specific ubiquitin carrier proteins contained a single functional MSA-like motif, which specifically controlled the expression of a reporter gene in the G2/M phase in BY2 cells. Furthermore, a genomic footprint experiment showed that the MSA element in the NtE2C promoter interacted with nuclear proteins in vivo. Therefore, we propose that the transcription of many G2/M phase-specific genes in tobacco is positively regulated by NtmybA2, in most cases through direct binding to the MSA elements.






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