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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Tnsd

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. This work unveils how Tn7 and Tn7-like elements determine the strict spacing between the target and integration sites. This is a preview of subscription content, access via your institution.

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A schematic representation of the attTn7 at the C-terminus of the glmS gene with the TnsD binding site is shown. New customer? The isolation of the TnsD truncation sequence 1 to ; full-length TnsD is aa as a dominant negative is consistent with this hypothesis see below. The transposition frequency of a miniTn 7 kan R element from the integration- and replication-defective phage KK1 into the chromosomal attTn7 site of E. As determined by analysis of a series of C-terminal deletion derivatives, the amino-terminal amino acid sequence 1 to of TnsD retain full binding to TnsC in this assay and thus contains the determinants for TnsD binding to TnsC. We also individually mutated Cys, Cys, Cys and His to serine, and analyzed the ability of these mutants to bind attTn7 in vitro and to promote transposition in vivo Table 2. Nucleic Acids Res. Your cart is empty. About this article Cite this article Mitra, R. BD is a derivative of E. Fields S, Song O: A novel genetic system to detect protein-protein interactions. Share this product. If ant is expressed, the phage grows lytically, whereas if expression of ant is blocked, lysogeny can be established. A The E. Phosphates are marked for clarity, although binding is probably dependent on base interactions, not sugar-backbone interactions.

Federal government websites often end in. The site is secure. 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.

Remembered your password? In another approach to identify the functional regions of TnsD, we conducted a screen for mutations in TnsD that have dominant-negative effects on TnsD-dependent transposition. To determine directly whether TnsD binding to attTn7 is dependent on zinc, gel-shift assays were performed using wild-type TnsD and attTn7 in the presence of increasing amounts of the zinc chelator 1,phenanthroline. Damages and issues Please inspect your order upon reception and contact us immediately if the item is defective, damaged or if you receive the wrong item, so that we can evaluate the issue and make it right. For the challenge phage experiment, we constructed a P22 phage with the E. Figure 4. In this study, we have made considerable progress in understanding the unique TnsD family of DNA-binding proteins by defining important protein determinants for DNA binding and transposition, and key nucleotides for the TnsD- attTn7 interaction. However, these truncated proteins did retain the ability to interact with TnsC, indicating that they were not simply unfolded we discuss interaction of TnsD with TnsC below. Google Scholar. Reasoning that TnsD would contact with the most highly conserved bases within this region, we mutated each of the most conserved bases in the E. Create your account Lost password? Download PDF. The activity of mutant attTn7 containing plasmid as a target for attTn7 transposition was measured using the lambda hop assay. Certain types of items cannot be returned, like custom products Such as video walls , and some audio products.