ISZm1068: an IS5-like insertion element from Zymomonas mobilis

A new insertion sequence, designated ISZm1068, was isolated from Zymomonas mobilis strain CP4. This element consists of 1,068 bp and contains one major ORF which shows similarities both at the nucleotide and at the amino acid sequence level with the corresponding ORFs encoding the transposases of many IS5 family elements, in particular the IS1031 group. Moreover, the Z. mobilis ORF shares the conserved N2, N3 and C1 signature motifs of the IS4 and IS5 families. Six out of seven Z. mobilis wild-type strains were shown by hybridisation to contain a single copy of the ISZm1068 element. Nucleotide sequences of the insertion elements from these strains exhibited extremely high levels of identity, varying from 94.25 to 99.25%. ISZm1068 was shown to be active in Escherichia coli cells and led to plasmid replicon fusions within the host cell. Sequence analysis of rare cointegration and resolution derivatives suggests that ISZm1068 has putative imperfect inverted repeats at its extremities of 18 bp (IR-right) and 14 bp (IR-left), and that a 3-bp (5'-TCA-3') target sequence is duplicated upon insertion.     

A widespread occurrence of extra open reading frames in plant Ty3/gypsy retrotransposons

Long terminal repeat (LTR) retrotransposons make up substantial parts of most higher plant genomes where they accumulate due to their replicative mode of transposition. Although the transposition is facilitated by proteins encoded within the gag-pol region which is common to all autonomous elements, some LTR retrotransposons were found to potentially carry an additional protein coding capacity represented by extra open reading frames located upstream or downstream of gag-pol. In this study, we performed a comprehensive in silico survey and comparative analysis of these extra open reading frames (ORFs) in the group of Ty3/gypsy LTR retrotransposons as the first step towards our understanding of their origin and function. We found that extra ORFs occur in all three major lineages of plant Ty3/gypsy elements, being the most frequent in the Tat lineage where most (77?%) of identified elements contained extra ORFs. This lineage was also characterized by the highest diversity of extra ORF arrangement (position and orientation) within the elements. On the other hand, all of these ORFs could be classified into only two broad groups based on their mutual similarities or the presence of short conserved motifs in their inferred protein sequences. In the Athila lineage, the extra ORFs were confined to the element 3' regions but they displayed much higher sequence diversity compared to those found in Tat. In the lineage of Chromoviruses the extra ORFs were relatively rare, occurring only in 5' regions of a group of elements present in a single plant family (Poaceae). In all three lineages, most extra ORFs lacked sequence similarities to characterized gene sequences or functional protein domains, except for two Athila-like elements with similarities to LOGL4 gene and part of the Chromoviruses extra ORFs that displayed partial similarity to histone H3 gene. Thus, in these cases the extra ORFs most likely originated by transduction or recombination of cellular gene sequences. In addition, the protein domain which is otherwise associated with DNA transposons have been detected in part of the Tat-like extra ORFs, pointing to their origin from an insertion event of a mobile element.     

A new MITE family,Pangrangja, in Gramineae species

The miniature inverted-repeat transposable element (MITE) is a type of class II transposon. We have isolated a new MITE, Pangrangja, from the species Oryza. Although Pangrangja elements are present in a number of Gramineae species, they are more prevalent in A genome Oryza species, Triticum, Aegilops, Hordeum, and Tripsacum. The Pangrangja has a 16 bp terminal inverted repeat (TIR) and conserved trinucleotides 5'-TTG-3' and 5'-AAA-3' at both ends. The TIR starts with 5'-CAGT-3', similar to the MITEs of the Emigrant family. The sequences between TIRs are very AT-rich and vary in length from 161 bp in A genome Oryza species to 513 bp in Hordeum vulgare. While the size and sequence of Pangrangia elements are conserved in the A genome Oryza species, there is some sequence variation in other plants. In an analysis of the mobile history of Oryza sativa, eight RESites (related to empty sites) were found and grouped into three types depending on the presence or absence of the conserved trinucleotides 5'-TTG-3' and 5'-AAA-3'. Since one of the RESites showed a perfect palindromic 22 bp sequence into which 180 bp of the Pangrangja element was inserted, the possible role of secondary structure of the palindromic sequences is discussed. We also suggest that as Pangrangja MITE-AFLP has been successful in many different Gramineae, Pangrangja elements may be useful in the genomic analysis of grasses.