Tnsd

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Metrics details. The bacterial transposon Tn7 is distinguished by its ability to recognize a specific site called attTn7 , and insert just downstream of the highly conserved chromosomal glmS gene. In this study, we identified important nucleotides within this region that are crucial for TnsD- attTn7 interaction. We also probed the regions of TnsD that interact with attTn7 and found that there are important DNA-binding determinants throughout the entire length of the protein, including an amino-terminal CCCH zinc-finger motif. Strikingly, whereas most transposons insert relatively randomly into many different sites, Tn 7 transposition is quite specific.

Tnsd

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Using the challenge phage assay, we also evaluated the ability of most of the dominant-negative TnsD missense mutants to bind to tnsd in vivoand found that all were significantly reduced in their ability to bind attTn7 Table 3, tnsd. Vidal M, tnsd, Legrain P: Yeast forward and reverse 'n'-hybrid systems.

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Thank you for visiting nature. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser or turn off compatibility mode in Internet Explorer. In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript. Tn7 transposable elements are unique for their highly specific, and sometimes programmable, target-site selection mechanisms and precise insertions. Owing to its multiple functions, TnsC is considered the linchpin in the Tn7 element. TnsC forms an asymmetric ring on target DNA that segregates target-site selection and interaction with the paired-end complex to opposite faces of the ring. By recognizing pre-distorted substrates, TnsC creates a built-in regulatory mechanism where ATP hydrolysis abolishes ring formation proximal to an existing element.

Tnsd

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Sorry, a shareable link is not currently available for this article. We also examined the ability of the dominant-negative TnsD mutants to promote transposition in the presence of TnsABC alone to determine if they possess residual TnsD activity. All had substantial defects in TnsD activity, with reductions ranging from at least 2. Craig N: Tn7. Biochem Biophys Acta. TnsD is a unique, sequence-specific, DNA-binding protein. Thus, P22 lysogeny can be a measure of protein binding; lysogeny reflects binding of TnsD, whereas if TnsD does not bind, ant is expressed and the phage grows lytically. Not surprisingly for a region that encodes a highly conserved protein, the first two positions at each codon are highly conserved, and variation is seen only at the third 'wobble' position. Google Scholar. Now there are many examples in GenBank of Tn7 relatives in a variety of different bacteria that contain obvious TnsD homologs Figure 4 , Figure 5 , Figure 6. Thus the challenge phage assay can be successfully used to evaluate TnsD binding to attTn7 in vivo. Maloy and J. The 3. DNA-binding reactions Binding reactions were performed with wild-type and mutant versions of TnsD proteins and wild-type TnsC protein as described previously [ 6 ]. Published : 23 July

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This fee is in place to cover the costs associated with restocking and processing the returned items. Add to cart. I n vitro transposition reactions were performed essentially as described previously [ 19 ]. The 36 bp TnsD-binding site lies within the 3' end of glmS , which encodes the C-terminus of the protein. Transformants were plated onto MacConkey lactose plates containing appropriate antibiotics. This region includes the GlmS active site and is highly conserved at the protein level. Crosslinking between TnsD and attTn7. Metrics details. A suspension system in its single roller design features 3 or 4 thin cables eliminating the need for black drop — the black border above the projection screen. Received : 07 May Challenge phages were constructed as described in [ 12 ]. EMBO J.