Kapler G M, Orias E, Blackburn E H
Department of Microbiology and Immunology, University of California, San Francisco 94143.
Genetics. 1994 Jun;137(2):455-66. doi: 10.1093/genetics/137.2.455.
The abundant rDNA minichromosome of Tetrahymena thermophila is generated by a series of developmentally controlled processing steps, termed rDNA maturation, during the formation of the new macronucleus in conjugating cells. rDNA maturation involves excision of a region encoding the single copy rRNA gene (rDNA) from its germline location, rearrangement of the rDNA into a palindromic minichromosome, de novo telomere addition, and amplification to approximately 10(4) copies. The rDNA is maintained at this high level in vegetatively growing cells. Using a previously developed genetic scheme for studying rDNA maturation and maintenance, we report the isolation of a new class of mutants defective for rDNA maturation. Several new rDNA maintenance mutants were also obtained. The maturation mutant, rmm10, is severely defective for the production of both monomeric and palindromic rDNA in the developing macronucleus. The mm10 mutation is recessive-lethal and cis-acting. None of the previously identified DNA sequence elements that control rDNA maturation or maintenance is mutated in rmm10. Therefore, additional cis-acting sequence elements must be required for rDNA maturation. Based on our current understanding of rDNA maturation processes, we suggest that the rmm10 mutation affects rDNA excision rather than subsequent rDNA amplification/replication.
嗜热四膜虫丰富的核糖体DNA(rDNA)微型染色体是在接合细胞形成新大核的过程中,通过一系列受发育控制的加工步骤产生的,这些步骤被称为rDNA成熟。rDNA成熟包括从其种系位置切除编码单拷贝rRNA基因(rDNA)的区域、将rDNA重排为回文微型染色体、从头添加端粒以及扩增至约10⁴个拷贝。rDNA在营养生长的细胞中维持在这一高水平。利用先前开发的用于研究rDNA成熟和维持的遗传方案,我们报告了一类新的rDNA成熟缺陷突变体的分离。还获得了几个新的rDNA维持突变体。成熟突变体rmm10在发育中的大核中产生单体和回文rDNA方面存在严重缺陷。mm10突变是隐性致死且顺式作用的。在rmm10中,先前鉴定的控制rDNA成熟或维持的DNA序列元件均未发生突变。因此,rDNA成熟必定需要其他顺式作用序列元件。基于我们目前对rDNA成熟过程的理解,我们认为rmm10突变影响rDNA切除而非随后的rDNA扩增/复制。