Overexpression of NtmybA2 in tobacco BY-2 cells
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ABSTRACT:
ORGANISM(S): Nicotiana tabacum
PROVIDER: GSE14251 | GEO | 2009/02/28
SECONDARY ACCESSION(S): PRJNA111181
REPOSITORIES: GEO
Similar Datasets
Project description:Promoters of 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. A least three R1R2R3-Myb proteins are expressed in tobacco (Nicotiana tabacum). To examine the genome-wide effects of NtmybA2 overexpression, we constructed transgenic tobacco BY-2 cell lines overexpressing full-length NtmybA2 protein. In addition, we generated transgenic BY-2 cell lines that overexpress truncated hyperactive form of NtmybA2 (NtmybA2âC) lacking its C-terminal region which negatively regulates its own activity for transcriptional activation. We used a custom-made 16K cDNA microarray for comparative transcriptome analysis of these transgenic tobacco BY-2 cell lines. As control sample, BY-2 cells transformed with empty vector pPZP-35S were used. The transgenic cell lines were analyzed during stationary phase (i.e. day 7-8 after subculture into fresh media). Keywords: Genetic modification 1. pPZP-35S vector control, 2. NtmybA2 overexpressor, 3. C-truncated NtmybA2 (NtmyA2âC) overexpressor
2010-05-22 | E-GEOD-14251 | biostudies-arrayexpress
Project description:Arabidopsis genome has 5 genes that encode for R1R2R3-type Myb transcription factors. We showed previously that two R1R2R3-Myb transcriptional factors, MYB3R1 and MYB3R4, bind to cis-acting motifs called MSA, which are commonly present in many G2/M-specific genes and act as transcriptional activator. Here, we performed genome-wide approach to identify target genes of MYB3R3, which, together with MYB3R1 and MYB3R5, have potential functions in transcriptional repression of G2/M-specific genes. Our ChIP-seq analysis revealed that MYB3R3 preferentially binds to the G2/M-specific genes in vivo. In addition, our data also showed that MYB3R3 associates with the predicted target genes of E2F, well known transcription factors acting during G1/S. These results suggested that MYB3R3 may not only directly bind G2/M-specific genes though recognition of the MSA elements, but also indirectly associates with DNA replication genes through physical interaction with E2F.
2015-06-01 | GSE60554 | GEO
Project description:R1R2R3-Myb proteins represent an evolutionarily conserved class of Myb family proteins important for cell cycle regulation and differentiation in eukaryotic cells. In plants, this class of Myb proteins are believed to play important roles in cell cycle regulation through transcriptional regulation of G2/M phase-specific genes by binding to common cis-elements, called MSA elements. In Arabidopsis thaliana, MYB3R1 and MYB3R4 act as transcriptional activators and positively regulate cytokinesis by activating transcription of KNOLLE, which encodes a cytokinesis-specific syntaxin. Here, we show that the double mutation myb3r1 myb3r4 causes pleiotropic developmental defects, some of which are due to deficiency of KNOLLE whereas other are not, suggesting multiple target genes are involved. Consistently, microarray analysis of the double mutant revealed altered expression of many genes, among which G2/M-specific genes showed significant overrepresentation of the MSA motif and a strong tendency to be down-regulated by the double mutation. Our results demonstrate, on a genome-wide level, the importance of the MYB3R-MSA pathway for regulating G2/M-specific transcription. In addition, MYB3R1 and MYB3R4 may have diverse roles during plant development by regulating G2/M-specific genes with various functions, as well as genes possibly unrelated to the cell cycle. Comparison between the myb3r1 myb3r4 double mutant and wild type in three paired replicates.
2011-09-06 | E-GEOD-29433 | biostudies-arrayexpress
Project description:R1R2R3-Myb proteins represent an evolutionarily conserved class of Myb family proteins important for cell cycle regulation and differentiation in eukaryotic cells. In plants, this class of Myb proteins are believed to play important roles in cell cycle regulation through transcriptional regulation of G2/M phase-specific genes by binding to common cis-elements, called MSA elements. In Arabidopsis thaliana, MYB3R1 and MYB3R4 act as transcriptional activators and positively regulate cytokinesis by activating transcription of KNOLLE, which encodes a cytokinesis-specific syntaxin. Here, we show that the double mutation myb3r1 myb3r4 causes pleiotropic developmental defects, some of which are due to deficiency of KNOLLE whereas other are not, suggesting multiple target genes are involved. Consistently, microarray analysis of the double mutant revealed altered expression of many genes, among which G2/M-specific genes showed significant overrepresentation of the MSA motif and a strong tendency to be down-regulated by the double mutation. Our results demonstrate, on a genome-wide level, the importance of the MYB3R-MSA pathway for regulating G2/M-specific transcription. In addition, MYB3R1 and MYB3R4 may have diverse roles during plant development by regulating G2/M-specific genes with various functions, as well as genes possibly unrelated to the cell cycle.
2011-09-07 | GSE29433 | GEO
Project description:Overexpression lines for each of NtMYBGR1 (Genbank accession No. AB279612) and NtDC1B (Genbank accession No. AB279614) were used to characterize the function of NtMYBGR1 and NtDC1B in tobacco BY-2 cells. We then analyzed the effect of overexpression of these genes on the expression of other genes, using the tobacco microarray. As control sample, tobacco cells transformed with empty vector pMAT137 vector were used. Two control and two transgenic cell lines were analyzed at day 4 after subculture into fresh media. Keywords: Genetic modification
2007-10-16 | GSE6949 | GEO
Project description:Temporal and spatial regulation of cell division is central for generating multicellular organs with predictable sizes and shapes. However, it remains largely unclear how genes with mitotic functions are transcriptionally regulated during organogenesis in plants. Here, we showed that a group of R1R2R3-Myb transcription factors are responsible for developmentally controlled downregulation of variety of mitotic genes in Arabidopsis. Loss of their functions resulted in elevated expression of mitotic genes in quiescent cells including those underwent terminal differentiation. Concomitantly, their mutations enhanced cell division activities in various aspects of plant development, generating organs with increased sizes and irregular architectures. In addition, we showed that this type of R1R2R3-Myb proteins are required for oscillated expression of G2/M-specfiic genes, most likely by inhibiting transcription outside of G2/M in the cell cycle. Our finding uncovered a novel plant-specific mechanism in which scheduled expression of G2/M-specific genes may require their global repression both in the cell cycle and during development.
2015-03-31 | GSE52298 | GEO
Project description:Temporal and spatial regulation of cell division is central for generating multicellular organs with predictable sizes and shapes. However, it remains largely unclear how genes with mitotic functions are transcriptionally regulated during organogenesis in plants. Here, we showed that a group of R1R2R3-Myb transcription factors are responsible for developmentally controlled downregulation of variety of mitotic genes in Arabidopsis. Loss of their functions resulted in elevated expression of mitotic genes in quiescent cells including those underwent terminal differentiation. Concomitantly, their mutations enhanced cell division activities in various aspects of plant development, generating organs with increased sizes and irregular architectures. In addition, we showed that this type of R1R2R3-Myb proteins are required for oscillated expression of G2/M-specfiic genes, most likely by inhibiting transcription outside of G2/M in the cell cycle. Our finding uncovered a novel plant-specific mechanism in which scheduled expression of G2/M-specific genes may require their global repression both in the cell cycle and during development. Seedlings (9d-old) or leaves (5d or 15d-old) of the wild type or myb3r mutants were sampled for RNA extraction and hybridization on Affymetrix microarrays.
2015-03-31 | E-GEOD-52298 | biostudies-arrayexpress
Project description:Overexpression lines for each of NtMYBGR1 (Genbank accession No. AB279612) and NtDC1B (Genbank accession No. AB279614) were used to characterize the function of NtMYBGR1 and NtDC1B in tobacco BY-2 cells. We then analyzed the effect of overexpression of these genes on the expression of other genes, using the tobacco microarray. As control sample, tobacco cells transformed with empty vector pMAT137 vector were used. Two control and two transgenic cell lines were analyzed at day 4 after subculture into fresh media. Keywords: Genetic modification 1. pMAT vector control, 2. NtMYBGR1 overexpressor, 3. NtDC1B overexpressor.
2007-10-15 | E-GEOD-6949 | biostudies-arrayexpress
Project description:We have previously reported that the responses of tobacco tissues treated with AaGlucan are similar to those for laminarin [1]. To better understand the overall responses and signaling pathways in tobacco, we have identified here AaGlucan and laminarin early response genes, using a custom 16K BY-2 microarray [2,3]. BY-2 cells were treated with laminarin or an AaGlucan-enriched fraction prepared from A. alternata 102 culture-medium and subjected to microarray analysis. [1] Shinya, T., Menard, R., Kozone, I., Matsuoka, H., Shibuya, N., Kauffmann, S., Matsuoka, K. and Saito, M. (2006) Novel b-1,3-, 1,6-oligoglucan elicitor from Alternaria alternata 102 for defense responses in tobacco. FEBS J. 273: 2421-2431. [2] Matsuoka, K., Demura, T., Galis, I., Horiguchi, T., Sasaki, M., Tashiro, G. and Fukuda, H. (2004) A comprehensive gene expression analysis toward the understanding of growth and differentiation of tobacco BY-2 cells. Plant Cell Physiol. 45: 1280-1289. [3] Galis, I., Simek, P., Narisawa, T., Sasaki, M., Horiguchi, T., Fukuda, H. and Matsuoka, K. (2006) A novel R2R3 MYB transcription factor NtMYBJS1 is a methyl jasmonate-dependent regulator of phenylpropanoid-conjugate biosynthesis in tobacco. Plant J. 46: 573-592. AaGlucan was purified from Alternaria alternata 102 as a potent elicitor for tobacco [1] Four-day-old tobacco BY-2 cells after subculture were treated with 10 microg/ml AaGlucan enriched fraction for 1h. Total RNA was prepared from the control and treated samples and used for microarray experiment after reverse transcription and labeling with Cy5 fluorescent dye. [1] Shinya, T., Menard, R., Kozone, I., Matsuoka, H., Shibuya, N., Kauffmann, S., Matsuoka, K. and Saito, M. (2006) Novel b-1,3-, 1,6-oligoglucan elicitor from Alternaria alternata 102 for defense responses in tobacco. FEBS J. 273: 2421-2431.
2007-10-16 | E-GEOD-6948 | biostudies-arrayexpress
Project description:We have previously reported that the responses of tobacco tissues treated with AaGlucan are similar to those for laminarin [1]. To better understand the overall responses and signaling pathways in tobacco, we have identified here AaGlucan and laminarin early response genes, using a custom 16K BY-2 microarray [2,3]. BY-2 cells were treated with laminarin or an AaGlucan-enriched fraction prepared from A. alternata 102 culture-medium and subjected to microarray analysis. [1] Shinya, T., Menard, R., Kozone, I., Matsuoka, H., Shibuya, N., Kauffmann, S., Matsuoka, K. and Saito, M. (2006) Novel b-1,3-, 1,6-oligoglucan elicitor from Alternaria alternata 102 for defense responses in tobacco. FEBS J. 273: 2421-2431. [2] Matsuoka, K., Demura, T., Galis, I., Horiguchi, T., Sasaki, M., Tashiro, G. and Fukuda, H. (2004) A comprehensive gene expression analysis toward the understanding of growth and differentiation of tobacco BY-2 cells. Plant Cell Physiol. 45: 1280-1289. [3] Galis, I., Simek, P., Narisawa, T., Sasaki, M., Horiguchi, T., Fukuda, H. and Matsuoka, K. (2006) A novel R2R3 MYB transcription factor NtMYBJS1 is a methyl jasmonate-dependent regulator of phenylpropanoid-conjugate biosynthesis in tobacco. Plant J. 46: 573-592. Keywords: Stress response
2007-10-16 | GSE6948 | GEO