笔记详情
标题
对EMSA方法学的讨论
内容
To validate the PBX2 and HOXA7 protein-protein interaction in SVOG cells, we examined their binding to two target sequences: PBX and EMX2. Previous studies showed that complexes of Pbx1 and HOX1–4 display optimal binding to the target sequence 5'-CGAATTGATTGATGCACTAATTGGAG-3' [23] and Pbx2 is also known to bind to this sequence [26]. TGAT is a Pbx binding site and TNAT is a HOX site. The TAAT, the HOX binding site which was used in this study, is accepted to bind the middle paralog groups 3–8 [29]. In addition, PBX2 and HOXA10 interactions with EMX2 were demonstrated in endometrial cancer cell lines [24]. EMX2 is expressed in the epithelial components of the urogenital system during development, and, as shown by its knockout studies, this gene is essential for the development of the female reproductive system [30]. In this study, HOXA7 and PBX2 complexes bound to the Pbx sequence, but not to the EMX2 sequence. The results indicate that a HOXA7 and PBX2 interaction occurs in granulosa cells. EMX2 can be a target of HOXA7, but it did not bind to PBX2 in granulosa cells. These results suggest that HOXA7 and PBX2 can make dimers in granulosa cells. However, when HOXA7 binds to the EMX2 promoter in granulosa cells, different cofactors might be used to enhance the HOXA7 binding specificity and strength.
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来源
Reproductive Biology and Endocrinology 2008, 6:49
类别
领域
Cancer/Oncology
