Lampson Bert C, Xu Chunying, Rice Scott A, Inouye Sumiko
Department of Health Sciences, East Tennessee State University, Johnson City, TN 37614, USA.
Gene. 2002 Oct 16;299(1-2):251-61. doi: 10.1016/s0378-1119(02)00977-0.
Retrons are reverse transcriptase (RT) encoding genetic elements usually located on the chromosome of a wide variety of mostly Gram-negative bacteria. Here we describe a new retron, designated Ne144, found in the chromosome of the myxobacterium Nannocystis exedens. This element codes for a 515-amino-acid RT that is most closely related to those found in other myxobacterial retrons. The RT is responsible for the production of a small satellite DNA called msDNA. This msDNA is composed of a 144 base, single-stranded DNA that is linked to a 72 base single-stranded RNA. The RNA strand is joined to the 5' end of the DNA chain via a 2'-5' linkage that occurs from the 2' position of an internal guanosine residue in the RNA. In addition to the retron element, the chromosome of N. exedens also contains several partial copies of the msDNA sequence as revealed by DNA hybridization experiments using msDNA as a probe. One of these partial copies was characterized from a chromosome restriction fragment and found to contain a sequence that matches the last 82 bases of the DNA strand and five bases of the RNA strand in msDNA-Ne144. This partial copy of msDNA is very likely a retrotransposed sequence that was generated by reverse transcription using an RNA (the primer-template RNA for msDNA) as a template and the 3' end of a nick in the chromosome as a primer, followed by incorporation into an open reading frame. The presence of this truncated copy of msDNA is strong evidence of retrotransposition in N. exedens causing an alteration in the bacterial genome.
反转录子是编码逆转录酶(RT)的遗传元件,通常位于多种主要为革兰氏阴性细菌的染色体上。在此,我们描述了一种新的反转录子,命名为Ne144,它存在于粘细菌嗜皮纳米囊菌的染色体中。该元件编码一种515个氨基酸的逆转录酶,与其他粘细菌反转录子中发现的逆转录酶最为相似。这种逆转录酶负责产生一种称为msDNA的小卫星DNA。这种msDNA由一个144个碱基的单链DNA组成,该单链DNA与一个72个碱基的单链RNA相连。RNA链通过2'-5'连接与DNA链的5'末端相连,这种连接发生在RNA内部鸟苷残基的2'位置。除了反转录子元件外,嗜皮纳米囊菌的染色体还包含msDNA序列的几个部分拷贝,这是通过使用msDNA作为探针的DNA杂交实验揭示的。其中一个部分拷贝是从染色体限制片段中鉴定出来的,发现其包含一个与msDNA-Ne144中DNA链的最后82个碱基和RNA链的5个碱基相匹配的序列。这种msDNA的部分拷贝很可能是一个反转录转座序列,它是通过以RNA(msDNA的引物模板RNA)为模板、以染色体上一个切口的3'末端为引物进行逆转录,然后整合到一个开放阅读框中产生的。这种截短的msDNA拷贝的存在是嗜皮纳米囊菌中发生反转录转座导致细菌基因组改变的有力证据。