301 Physical Science, Department of Botany, Oklahoma State University, Stillwater, OK, 74078, USA.
Plant Mol Biol. 2014 May;85(1-2):81-94. doi: 10.1007/s11103-013-0170-9. Epub 2014 Jan 16.
The Arabidopsis A1-type cyclin, CYCA1;2, also named TARDY ASYNCHRONOUS MEIOSIS (TAM), is known for its positive role in meiotic cell cycle progression, but its function in other cells has not been characterized. This paper reports the role of CYCA1;2/TAM in differentiated cells in vegetative organs. The pattern of CYCA1;2/TAM expression was investigated by promoter and protein fusions using the β-glucuronidase and the green fluorescent protein, respectively. The relevance of the promoter region used in these gene fusion constructs was verified by the effective complementation of the phenotype of the diploid null allele, tam-2 2C by a genomic fragment containing the wild-type coding region of CYCA1;2/TAM and the promoter region. CYCA1;2/TAM expression was found primarily in non-proliferating cells such as guard cells, trichomes, and mesophyll cells, and in vascular tissue. In two types of overexpression lines, one containing the CYCA1;2/TAM transgene driven by the ARABIDOPSIS SKP1-LIKE1 (ASK1) promoter and the other CYCA1;2/TAM-GFP driven by the cauliflower mosaic virus 35S promoter, the largest differences between the transgene transcript levels were approximately 72- and 45-folds, respectively, but the TAM-GFP signal levels in the mesophyll and stomata in the 35S:TAM-GFP lines only differ slightly. Furthermore, the GFP signals in the mesophyll and stomata in the TAM:TAM-GFP and 35S:TAM-GFP lines were all at similarly low levels. These results indicate that the CYCA1;2/TAM protein is likely maintained at low levels in these cells through post-transcriptional regulation. Loss of function in CYCA1;2/TAM resulted in increases in the nuclear size in both trichomes and guard cells. Surprisingly, overexpression of CYCA1;2/TAM led to similar increases. The large increases in trichome nuclear size likely reflected ploidy increases while the moderate increases in guard cell nuclear size did not justify for a ploidy increase. These nuclear size increases were not clearly correlated with trichome branch number increases and guard cell size increases, respectively. These results suggest that cellular homeostasis of the CYCA1;2/TAM protein is linked to the control of nuclear sizes in trichomes and guard cells.
拟南芥 A1 型细胞周期蛋白 CYCA1;2,也称为 TARDY ASYNCHRONOUS MEIOSIS (TAM),因其在减数分裂细胞周期进程中的积极作用而闻名,但它在其他细胞中的功能尚未得到表征。本文报道了 CYCA1;2/TAM 在营养器官的分化细胞中的作用。通过启动子和蛋白质融合分别使用β-葡萄糖醛酸酶和绿色荧光蛋白来研究 CYCA1;2/TAM 的表达模式。通过含有野生型 CYCA1;2/TAM 编码区和启动子区的基因组片段有效互补二倍体 null 等位基因 tam-2 2C 的表型,验证了这些基因融合构建物中使用的启动子区域的相关性。发现 CYCA1;2/TAM 表达主要存在于非增殖细胞中,如保卫细胞、毛状体和叶肉细胞以及维管束组织中。在两种过表达系中,一种含有由 ARABIDOPSIS SKP1-LIKE1 (ASK1) 启动子驱动的 CYCA1;2/TAM 转基因,另一种由花椰菜花叶病毒 35S 启动子驱动的 CYCA1;2/TAM-GFP,转基因转录物水平之间的最大差异分别约为 72 倍和 45 倍,但 35S:TAM-GFP 系中叶肉和气孔中的 TAM-GFP 信号水平仅略有差异。此外,TAM:TAM-GFP 和 35S:TAM-GFP 系中叶肉和气孔中的 GFP 信号均处于相似的低水平。这些结果表明,CYCA1;2/TAM 蛋白可能通过转录后调控在这些细胞中维持低水平。CYCA1;2/TAM 的功能丧失导致毛状体和保卫细胞中的核大小增加。令人惊讶的是,过表达 CYCA1;2/TAM 也导致类似的增加。毛状体核大小的大幅增加可能反映了倍性增加,而保卫细胞核大小的适度增加并不支持倍性增加。这些核大小的增加与毛状体分支数的增加和保卫细胞大小的增加分别没有明显的相关性。这些结果表明,CYCA1;2/TAM 蛋白的细胞内稳态与毛状体和保卫细胞中核大小的控制有关。