Graham K M, Ko C, Park K S, Sarge K, Park-Sarge O K
Department of Physiology, University of Kentucky, Lexington, Kentucky 40536-0084, USA.
Biochem Biophys Res Commun. 2000 Nov 11;278(1):48-57. doi: 10.1006/bbrc.2000.3759.
We have isolated a rat intracisternal-A particle element (IAP)-like element (IAP-LE) from ovarian granulosa cells that appears to be identical to the rat EST clone AA964260. The compiled cDNA sequences contain several putative in-frame translation initiation codons with the largest capable of encoding a 365 amino acid protein with a reverse transcriptase domain in the N-terminus as well as a bipartite nuclear localization signal sequence in the middle. Northern blotting shows a major approximately 7 Kb transcript and a minor approximately 5 Kb transcript that are abundantly expressed in the ovary. In situ hybridization histochemistry using ovaries from gonadotropin-treated immature rats and regularly cycling adult rats show that this transcript is predominantly localized to granulosa cells of all healthy follicles, including primary follicles, and to newly-formed and healthy corpora lutea. This cell-specific expression pattern of the IAP-LE gene is distinct from those of the several known retroviral elements, suggesting the potentially novel functional importance of the IAP-LE gene. Taken together, our results demonstrate abundant and cell-specific expression of a novel IAP-LE in rat granulosa cells.
我们从大鼠卵巢颗粒细胞中分离出一种大鼠脑池内A颗粒元件(IAP)样元件(IAP-LE),它似乎与大鼠表达序列标签(EST)克隆AA964260相同。编译后的cDNA序列包含几个推定的读框内翻译起始密码子,其中最大的能够编码一个365个氨基酸的蛋白质,该蛋白质在N端具有逆转录酶结构域,中间还有一个双分核定位信号序列。Northern印迹显示在卵巢中大量表达的主要约7kb转录本和次要约5kb转录本。使用促性腺激素处理的未成熟大鼠和正常周期成年大鼠的卵巢进行原位杂交组织化学显示,该转录本主要定位于所有健康卵泡(包括初级卵泡)的颗粒细胞以及新形成的健康黄体。IAP-LE基因的这种细胞特异性表达模式与几种已知逆转录病毒元件的表达模式不同,表明IAP-LE基因可能具有新的功能重要性。综上所述,我们的结果证明了一种新型IAP-LE在大鼠颗粒细胞中大量且细胞特异性表达。