Bailey A D, Pavelitz T, Weiner A M
Department of Molecular Biophysics, Yale University, New Haven, Connecticut 06520-8114, USA.
Mol Cell Biol. 1998 Apr;18(4):2262-71. doi: 10.1128/MCB.18.4.2262.
The multigene family encoding human U2 small nuclear RNA (snRNA) is organized as a single large tandem array containing 5 to 25 copies of a 6.1-kb repeat unit (the RNU2 locus). Remarkably, each of the repeat units within an individual U2 tandem array appears to be identical except for an irregular dinucleotide tract, known as the CT microsatellite, which exhibits minor length and sequence polymorphism. Using a somatic cell genetic assay, we previously noticed that the CT microsatellite appeared to stabilize artificial tandem arrays of U2 snRNA genes. We now demonstrate that the CT microsatellite is required to establish large tandem arrays of transcriptionally active U2 genes, increasing both the average and maximum size of the resulting arrays. In contrast, the CT microsatellite has no effect on the average or maximal size of artificial arrays containing transcriptionally inactive U2 genes that lack key promoter elements. Our data reinforce the connection between recombination and transcription. Active U2 transcription interferes with establishment or maintenance of the U2 tandem array, and the CT microsatellite opposes these effects, perhaps by binding GAGA or GAGA-related factors which alter local chromatin structure. We speculate that the mechanisms responsible for maintenance of tandem arrays containing active promoters may differ from those that maintain tandem arrays of transcriptionally inactive sequences.
编码人类U2小核RNA(snRNA)的多基因家族被组织成一个单一的大串联阵列,其中包含5到25个6.1 kb重复单元的拷贝(RNU2基因座)。值得注意的是,单个U2串联阵列中的每个重复单元似乎都是相同的,除了一个不规则的二核苷酸序列,即CT微卫星,它表现出较小的长度和序列多态性。使用体细胞遗传学检测方法,我们之前注意到CT微卫星似乎能稳定U2 snRNA基因的人工串联阵列。我们现在证明,CT微卫星是建立转录活性U2基因的大型串联阵列所必需的,它能增加所得阵列的平均大小和最大大小。相比之下,CT微卫星对包含缺乏关键启动子元件的转录无活性U2基因的人工阵列的平均大小或最大大小没有影响。我们的数据强化了重组与转录之间的联系。活跃的U2转录会干扰U2串联阵列的建立或维持,而CT微卫星可能通过结合GAGA或GAGA相关因子来改变局部染色质结构,从而对抗这些影响。我们推测,负责维持含有活跃启动子的串联阵列的机制可能与维持转录无活性序列的串联阵列的机制不同。