Specific nicking at the 3′ ends of the terminal inverted repeat sequences in transposon Tn3 by transposase and an E. coli protein ACP

Maekawa, Takafumi; Yanagihara, Keiko; Ohtsubo, Eiichi

doi:10.1046/j.1365-2443.1996.d01-221.x

Cited by 13 publications

(8 citation statements)

References 30 publications

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“…8). This is consistent with TnpA being a member of the DD-E/D transposase superfamily (221,222). However, the overall efficiency of the reaction was too low for the molecular characterization of transposition intermediates and products (220)(221)(222).…”

Section: Catalysis Of the Transposition Reactionssupporting

confidence: 71%

“…Our understanding of the molecular transposition mechanism of Tn3 family transposons has long remained scant because of the technical difficulties inherent in transposase purification for biochemical characterization. An in vitro transposition reaction based on TnpAenriched cell extracts was developed for Tn3 and, as expected, TnpA was shown to introduce specific nicks at the 3′ ends of the transposon in an Mg 2+ -dependent manner (220)(221)(222) (Fig. 8).…”

Section: Catalysis Of the Transposition Reactionsmentioning

confidence: 68%

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The Tn 3 -family of Replicative Transposons

Lambin²,

et al. 2015

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Transposons of the Tn3 family form a widespread and remarkably homogeneous group of bacterial transposable elements in terms of transposition functions and an extremely versatile system for mediating gene reassortment and genomic plasticity owing to their modular organization. They have made major contributions to antimicrobial drug resistance dissemination or to endowing environmental bacteria with novel catabolic capacities. Here, we discuss the dynamic aspects inherent to the diversity and mosaic structure of Tn3-family transposons and their derivatives. We also provide an overview of current knowledge of the replicative transposition mechanism of the family, emphasizing most recent work aimed at understanding this mechanism at the biochemical level. Previous and recent data are put in perspective with those obtained for other transposable elements to build up a tentative model linking the activities of the Tn3-family transposase protein with the cellular process of DNA replication, suggesting new lines for further investigation. Finally, we summarize our current view of the DNA site-specific recombination mechanisms responsible for converting replicative transposition intermediates into final products, comparing paradigm systems using a serine recombinase with more recently characterized systems that use a tyrosine recombinase.

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Section: Catalysis Of the Transposition Reactionssupporting

confidence: 71%

Section: Catalysis Of the Transposition Reactionsmentioning

confidence: 68%

The Tn 3 -family of Replicative Transposons

Lambin²,

et al. 2015

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“…Thus, it may be that our standard assay conditions are not conducive for studying the effects of host proteins on Tn7 transposition in vitro; it is possible that more dramatic effects can be observed under other assay conditions. ACP has also been reported to stimulate nicking at the 3Ј ends of Tn3 by the purified Tn3 transposase, although it is not clear whether ACP alters the binding of the Tn3 transposase to the 3Ј ends (Maekawa et al, 1996). Here we have been able to show a direct effect of ACP on Tn7 transposition in vitro.…”

Section: L29 and Acp Can Stimulate Tn7 Transpositionsupporting

confidence: 45%

“…ACP has also been reported to stimulate nicking at the 3′ ends of Tn 3 by the purified Tn 3 transposase, although it is not clear whether ACP alters the binding of the Tn 3 transposase to the 3′ ends (Maekawa et al ., 1996). Here we have been able to show a direct effect of ACP on Tn 7 transposition in vitro .…”

Section: Discussionmentioning

confidence: 99%

Host proteins can stimulate Tn7 transposition: a novel role for the ribosomal protein L29 and the acyl carrier protein

Sharpe

Craig

1998

The EMBO Journal

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The bacterial transposon Tn7 is distinguished by its ability to insert at a high frequency into a specific site in the Escherichia coli chromosome called attTn7. Tn7 insertion into attTn7 requires four Tn7-encoded transposition proteins: TnsA, TnsB, TnsC and TnsD. The selection of attTn7 is determined by TnsD, a sequence-specific DNA-binding protein. TnsD binds attTn7 and interacts with TnsABC, the core transposition machinery, which facilitates the insertion of Tn7 into attTn7. In this work, we report the identification of two host proteins, the ribosomal protein L29 and the acyl carrier protein (ACP), which together stimulate the binding of TnsD to attTn7. The combination of L29 and ACP also stimulates Tn7 transposition in vitro. Interestingly, mutations in L29 drastically decrease Tn7 transposition in vivo, and this effect of L29 on Tn7 transposition is specific for TnsABC⍣D reactions.

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“…Some of these diverse proteins were MukB, a protein involved in the partition of chromosomes between daughter cells; SecA, a key protein translocation component; and SpoT, a protein required for the adaptation of cell physiology to nutrient limitation. Moreover, ACP had previously been reported to be an essential cofactor for the synthesis of a periplasmic oligosaccharide in vitro (49,50), to stimulate the nicking reaction of transposon Tn3 (26), and to give improved binding of Tn7 transposase to its target sequence (44).…”

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confidence: 99%

Gene-Specific Random Mutagenesis of Escherichia coli In Vivo: Isolation of Temperature-Sensitive Mutations in the Acyl Carrier Protein of Fatty Acid Synthesis

Lay¹,

Cronan

2006

J Bacteriol

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Acyl carrier proteins (ACPs) are very small acidic proteins that play a key role in fatty acid and complex lipid synthesis. Moreover, recent data indicate that the acyl carrier protein of Escherichia coli has a large protein interaction network that extends beyond lipid synthesis. Despite extensive efforts over many years, no temperature-sensitive mutants with mutations in the structural gene (acpP) that encodes ACP have been isolated. We report the isolation of three such mutants by a new approach that utilizes error-prone PCR mutagenesis, overlap extension PCR, and phage Red-mediated homologous recombination and that should be generally applicable. These mutants plus other experiments demonstrate that ACP function is essential for the growth of E. coli. Each of the mutants was efficiently modified with the phosphopantetheinyl moiety essential for the function of ACP in lipid synthesis, and thus lack of function at the nonpermissive temperature cannot be attributed to a lack of prosthetic group attachment. All of the mutant proteins were largely stable at the nonpermissive temperature except the A68T/N73D mutant protein. Fatty acid synthesis in strains that carried the D38V or A68T/N73D mutations was inhibited upon a shift to the nonpermissive temperature and in the latter case declined to a small percentage of the rate of the wild-type strain.Two systems for fatty acid biosynthesis, called types I and II, are found in nature. Type I systems occur in the cytosols of mammalian and plant cells, in some plant plastids, and in a few bacteria. In these systems, the steps of fatty acid biosynthesis are catalyzed by one or two very large polypeptides that contain multiple active sites (47). In contrast, the type II systems which occur in bacteria, mitochondria, most plant plastids, and protozoan apicoplasts produce fatty acids using a series of soluble enzymes in which each protein has a discrete enzymatic activity (38). A key component of both systems is acyl carrier protein (ACP) which constitutes a domain of the type I multifunctional proteins but is a very small, soluble, and highly acidic protein in type II systems. ACP is functional only in fatty acid biosynthesis after it has been posttranslationally modified by covalent attachment of a 4Ј-phosphopantetheinyl (4Ј-PP) moiety. The 4Ј-PP prosthetic group is attached to the hydroxyl group of a centrally located serine residue by the AcpS 4Ј-PP transferase. Acyl intermediates are bound to the 4Ј-PP thiol in a thioester linkage that allows ACP to shuttle intermediates among the fatty acid synthetic enzymes. The thioester linkage also serves to facilitate chemistry at acyl chain ␤-carbons.Escherichia coli contains a single ACP encoded by the acpP gene (35). This was the first ACP discovered (40, 52) and remains the best studied of these proteins. Despite the breadth and depth of studies of E. coli ACP and its acylated derivatives, no mutant strain encoding a mutant ACP has been reported. However, ACP seemed certain to be an essential protein since mutants in several f...

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**Specific nicking at the 3′ ends of the terminal inverted repeat sequences in transposon Tn3 by transposase and an E. coli protein ACP**

Cited by 13 publications

References 30 publications

The Tn 3 -family of Replicative Transposons

The Tn 3 -family of Replicative Transposons

Host proteins can stimulate Tn7 transposition: a novel role for the ribosomal protein L29 and the acyl carrier protein

Gene-Specific Random Mutagenesis of Escherichia coli In Vivo: Isolation of Temperature-Sensitive Mutations in the Acyl Carrier Protein of Fatty Acid Synthesis

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