Mullin D A, Zies D L, Mullin A H, Caballera N, Ely B
Department of Cell and Molecular Biology, Tulane University, New Orleans, LA 70118, USA.
Mol Gen Genet. 1997 Apr 28;254(4):456-63. doi: 10.1007/s004380050439.
IS511 is an endogenous insertion sequence (IS) of the bacterium Caulobacter crescentus strain CB15 and it is the first Caulobacter IS to be characterized at the molecular level. We determined the 1266-bp nucleotide sequence of IS511 and investigated its genetic organization, relationship to other ISs, and transposition properties. IS511 belongs to a distinct branch of the IS3 family that includes ISR1, IS476, and IS1222, based on nucleotide sequence similarity. The nucleotide sequence of IS511 encodes open reading frames (orfs) designated here as orfA and orfB, and their relative organization and amino acid sequences of the predicted protein products are very similar to those of orfAs and orfBs of other IS3 family members. Nuclease S1 protection assays identified an IS511 RNA, and its 5' end maps approximately 16 nucleotides upstream of orfA and about six nucleotides downstream of a sequence that is similar to the consensus sequence of C. crescentus housekeeping promoters. Evidence is presented that IS511 is capable of precise excision from the chromosome, and transposition from the chromosome to a plasmid. Transpositional insertions of IS511 occurred within sequences with a relatively high G + C content, and they were usually, but not always, flanked by a 4-bp direct repeat that matches a sequence at the site of insertion. We also determined the nucleotide sequence flanking the four endogenous IS511 elements that reside in the chromosome of C. crescentus. Our findings demonstrate that IS511 is a transposable IS that belongs to a branch of the IS3 family.
IS511是新月柄杆菌CB15菌株的一种内源性插入序列(IS),它是第一个在分子水平上得到表征的柄杆菌IS。我们测定了IS511的1266个碱基对的核苷酸序列,并研究了其基因组织、与其他IS的关系以及转座特性。基于核苷酸序列相似性,IS511属于IS3家族的一个独特分支,该分支包括ISR1、IS476和IS1222。IS511的核苷酸序列编码在此指定为orfA和orfB的开放阅读框(orf),它们的相对组织以及预测的蛋白质产物的氨基酸序列与其他IS3家族成员的orfA和orfB非常相似。核酸酶S1保护试验鉴定出一种IS511 RNA,其5'端位于orfA上游约16个核苷酸处,以及一个与新月柄杆菌管家启动子的共有序列相似的序列下游约6个核苷酸处。有证据表明,IS511能够从染色体上精确切除,并从染色体转座到质粒上。IS511的转座插入发生在G + C含量相对较高的序列中,它们通常(但并非总是)两侧有一个4碱基对的直接重复序列,该序列与插入位点的序列匹配。我们还测定了位于新月柄杆菌染色体上的四个内源性IS511元件侧翼的核苷酸序列。我们的研究结果表明,IS511是一种可转座的IS,属于IS3家族的一个分支。