van Leeuwen F, Taylor M C, Mondragon A, Moreau H, Gibson W, Kieft R, Borst P
Division of Molecular Biology, The Netherlands Cancer Institute, Plesmanlaan 121, 1066 CX Amsterdam, The Netherlands.
Proc Natl Acad Sci U S A. 1998 Mar 3;95(5):2366-71. doi: 10.1073/pnas.95.5.2366.
The unusual DNA base beta-D-glucosyl-hydroxymethyluracil, called "J, " replaces approximately 0.5-1% of Thy in DNA of African trypanosomes but has not been found in other organisms thus far. In Trypanosoma brucei, J is located predominantly in repetitive DNA, and its presence correlates with the silencing of telomeric genes. Using antibodies specific for J, we have developed sensitive assays to screen for J in a range of organisms and have found that J is not limited to trypanosomes that undergo antigenic variation but is conserved among Kinetoplastida. In all kinetoplastids tested, including the human pathogens Leishmania donovani and Trypanosoma cruzi, J was found to be abundantly present in the (GGGTTA)n telomere repeats. Outside Kinetoplastida, J was found only in Diplonema, a small phagotrophic marine flagellate, in which we also identified 5-MeCyt. Fractionation of Diplonema DNA showed that the two modifications are present in a common genome compartment, which suggests that they may have a similar function. Dinoflagellates appear to contain small amounts of modified bases that may be analogs of J. The evolutionary conservation of J in kinetoplastid protozoans suggests that it has a general function, repression of transcription or recombination, or a combination of both. T. brucei may have recruited J for the control of genes involved in antigenic variation.
异常的DNA碱基β-D-葡糖基羟甲基尿嘧啶,称为“J”,在非洲锥虫的DNA中取代了约0.5 - 1%的胸腺嘧啶(Thy),但迄今为止在其他生物中尚未发现。在布氏锥虫中,J主要位于重复DNA中,其存在与端粒基因的沉默相关。利用针对J的特异性抗体,我们开发了灵敏的检测方法来筛选一系列生物中的J,发现J并不局限于经历抗原变异的锥虫,而是在动质体目生物中保守存在。在所有测试的动质体生物中,包括人类病原体杜氏利什曼原虫和克氏锥虫,都发现J大量存在于(GGGTTA)n端粒重复序列中。在动质体目之外,仅在双滴虫属(一种小型吞噬性海洋鞭毛虫)中发现了J,在该生物中我们还鉴定出了5-甲基胞嘧啶(5-MeCyt)。双滴虫属DNA的分级分离表明,这两种修饰存在于共同的基因组区域,这表明它们可能具有相似的功能。甲藻似乎含有少量可能是J类似物的修饰碱基。J在动质体原生动物中的进化保守性表明它具有一般功能,即抑制转录或重组,或两者兼而有之。布氏锥虫可能已招募J来控制参与抗原变异的基因。