Vazquez M, Ben-Dov C, Lorenzi H, Moore T, Schijman A, Levin M J
Laboratorio de Biología Molecular de la Enfermedad de Chagas, Instituto de Investigaciones en Ingenieria Genética y Biologia Molecular, Vuelta de Obligado 2490, 1428 Buenos Aires, Argentina.
Proc Natl Acad Sci U S A. 2000 Feb 29;97(5):2128-33. doi: 10.1073/pnas.050578397.
The short interspersed repetitive element (SIRE) of Trypanosoma cruzi was first detected when comparing the sequences of loci that encode the TcP2beta genes. It is present in about 1,500-3,000 copies per genome, depending on the strain, and it is distributed in all chromosomes. An initial analysis of SIRE sequences from 21 genomic fragments allowed us to derive a consensus nucleotide sequence and structure for the element, consisting of three regions (I, II, and III) each harboring distinctive features. Analysis of 158 transcribed SIREs demonstrates that the consensus is highly conserved. The sequences of 51 cDNAs show that SIRE is included in the 3' end of several mRNAs, always transcribed from the sense strand, contributing the polyadenylation site in 63% of the cases. This study led to the characterization of VIPER (vestigial interposed retroelement), a 2,326-bp-long unusual retroelement. VIPER's 5' end is formed by the first 182 bp of SIRE, whereas its 3' end is formed by the last 220 bp of the element. Both SIRE moieties are connected by a 1,924-bp-long fragment that carries a unique ORF encoding a complete reverse transcriptase-RNase H gene whose 15 C-terminal amino acids derive from codons specified by SIRE's region II. The amino acid sequence of VIPER's reverse transcriptase-RNase H shares significant homology to that of long terminal repeat retrotransposons. The fact that SIRE and VIPER sequences are found only in the T. cruzi genome may be of relevance for studies concerning the evolution and the genome flexibility of this protozoan parasite.
克氏锥虫的短散在重复元件(SIRE)最初是在比较编码TcP2β基因的基因座序列时被检测到的。根据菌株不同,每个基因组中大约有1500 - 3000个拷贝,并且分布在所有染色体上。对来自21个基因组片段的SIRE序列进行初步分析,使我们能够推导该元件的共有核苷酸序列和结构,其由三个区域(I、II和III)组成,每个区域都有独特特征。对158个转录的SIRE进行分析表明,该共有序列高度保守。51个cDNA的序列显示,SIRE包含在几个mRNA的3'端,总是从有义链转录,在63%的情况下提供多聚腺苷酸化位点。这项研究导致了对VIPER(残留插入反转录元件)的鉴定,它是一个2326 bp长的不寻常反转录元件。VIPER的5'端由SIRE的前182 bp形成,而其3'端由该元件的最后220 bp形成。两个SIRE部分通过一个1924 bp长的片段连接,该片段携带一个独特的开放阅读框,编码一个完整的逆转录酶 - RNase H基因,其15个C末端氨基酸来自SIRE区域II指定的密码子。VIPER的逆转录酶 - RNase H的氨基酸序列与长末端重复反转录转座子的氨基酸序列有显著同源性。SIRE和VIPER序列仅在克氏锥虫基因组中被发现这一事实,可能与关于这种原生动物寄生虫的进化和基因组灵活性的研究相关。