Wang Yixing, Yang Ming
Department of Botany, Oklahoma State University, 104 Life Sciences East, Stillwater, OK 74078, USA.
Planta. 2006 Feb;223(3):613-7. doi: 10.1007/s00425-005-0154-3. Epub 2005 Nov 9.
Normal progression of genetic recombination requires timely degradation of many proteins, but little is known about the proteolytic mechanism. The ARABIDOPSIS SKP1-LIKE1 (ASK1) protein is a component of the Skp1-Cullin-F-box-protein (SCF) ubiquitin ligases that target a variety of proteins for degradation via the 26S proteasome pathway. Previous studies indicate that the early defects of the mutant ask1-1 occur in a prophase-I period overlapping with the period of homologous recombination. We provide evidence in this report that ASK1 is predominately expressed from leptotene to pachytene, and negatively regulates recombination. First, the ASK1 transcript was found not to co-exist with that of its closest homolog ASK2 only during prophase I of male meiosis, suggesting that ASK1 is functionally non-redundant only in prophase I. Second, the peak level of an ASK1-green fluorescence protein (GFP) fusion protein expressed by an ASK1 promoter region occurred only from leptotene to pachytene. The ASK1-GFP in a dominant negative fashion resulted in abnormal tetrads resembling those of the ask1-1 mutant, supporting that the expression timing of the ASK1-GFP in male meiocytes reflects the expression timing of the endogenous ASK1. Lastly, using a marker for recombination events, a significant increase in recombination frequency was detected in plants heterozygous for ask1-1. These results indicate that ASK1 normally plays a repressive role in male recombination in Arabidopsis.
基因重组的正常进行需要许多蛋白质的及时降解,但关于蛋白水解机制却知之甚少。拟南芥类SKP1-1(ASK1)蛋白是Skp1- Cullin-F-盒蛋白(SCF)泛素连接酶的一个组分,该连接酶通过26S蛋白酶体途径靶向多种蛋白质进行降解。先前的研究表明,突变体ask1-1的早期缺陷发生在与同源重组时期重叠的减数分裂前期I。我们在本报告中提供证据表明,ASK1主要在细线期到粗线期表达,并对重组起负调控作用。首先,仅在雄配子减数分裂前期I发现ASK1转录本与其最接近的同源物ASK2的转录本不共存,这表明ASK1仅在减数分裂前期I功能上无冗余。其次,由ASK1启动子区域表达的ASK1-绿色荧光蛋白(GFP)融合蛋白的峰值水平仅出现在细线期到粗线期。以显性负性方式存在的ASK1-GFP导致出现类似于ask1-1突变体的异常四分体,这支持了雄配子细胞中ASK1-GFP的表达时间反映了内源性ASK1的表达时间。最后,使用重组事件标记,在ask1-1杂合植物中检测到重组频率显著增加。这些结果表明,ASK1在拟南芥雄配子重组中通常起抑制作用。