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Resources » Paper

Vos JC et al. (1992) Worm Breeder's Gazette "The Transposable Element Tc1 Encodes a Protein that Specifically Binds to Tc1 DNA Ends"

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  • Comments on Vos JC et al. (1992) Worm Breeder's Gazette "The Transposable Element Tc1 Encodes a Protein that Specifically Binds to Tc1 DNA Ends" (0)

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    Status:
    Publication type:
    Gazette_article
    WormBase ID:
    WBPaper00014602

    Vos JC, van Luenen HGAM, Groenen JTM, & Plasterk RHA (1992). The Transposable Element Tc1 Encodes a Protein that Specifically Binds to Tc1 DNA Ends. Worm Breeder's Gazette, 12(3), 87. Unpublished information; cite only with author permission.

    Originally a 273 triplet ORF has been identified in the first sequence of the transposable element Tc 1 of C. elegans (Rosenzweig et al., NAR 11: 4201, 1983). However, analysis of other Tc1 elements and a related element in C. briggsae suggested the presence of a longer ORF which would be interrupted by an intron (Schukkink and Plasterk, NAR 18: 895, 1990; Prasad et al., Genome 34: 6, 1991). Attempts to directly clarify the identity of the ORF by either RNA or protein analysis in C. elegans have been unsuccessful due to the very low level of expression. In order to increase the level of expression, we fused the most 5' ATG start codon in Tc1 to the hsp-16 promoter (Russnak et al., MCB 5: 1268, 1985). After establishing a transgenic worm with this construct, we analyzed the heat shock induced RNA. A 41 nt intron was removed from the primary transcripts for which the 3' splice site was as predicted previously [See Figure 1]. We conclude that Tc1 encodes a 343 amino acid polypeptide, the putative transposase, which we will refer to as Tc1A .Currently, we are investigating the expression of Tc 1 A in the transgenic worm and the effect on Tc1 transposition. We have expressed the 343 amino acid Tc1 A protein in E. coli. A soluble protein was partially purified and it could be shown by gel retardation assays that it binds in a sequence-specific fashion to the 55 bp inverted repeat of Tc1 .The binding properties of Tc1A will be further analyzed and the protein will be tested for its ability to act in an in vitro transposition assay either on its own or together with extracts prepared from C. elegans.


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