scholarly journals Role of Secondary Attachment Sites in Changing the Specificity of Site-Specific Recombination

2006 ◽  
Vol 188 (9) ◽  
pp. 3409-3411 ◽  
Author(s):  
Edit Rutkai ◽  
Andrea György ◽  
László Dorgai ◽  
Robert A. Weisberg
Keyword(s):  
Wild Type ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Attachment Sites ◽  
Core Site ◽  
Secondary Attachment ◽  
Lambdoid Phages

ABSTRACT We previously proposed that lambdoid phages change their insertion specificity by adapting their integrases to sequences found in secondary attachment sites. To test this model, we quantified recombination between partners that carried sequences from secondary attachment sites catalyzed by wild-type and by mutant integrases with altered specificities. The results are consistent with the model, and indicate differential core site usage in excision and integration.

scholarly journals Transfer of the Symbiotic Plasmid of Rhizobium etli CFN42 Requires Cointegration with p42a, Which May Be Mediated by Site-Specific Recombination

2004 ◽  
Vol 186 (22) ◽  
pp. 7538-7548 ◽  
Author(s):  
Susana Brom ◽  
Lourdes Girard ◽  
Cristina Tun-Garrido ◽  
Alejandro García-de los Santos ◽  
Patricia Bustos ◽  
...  
Keyword(s):  
Attachment Site ◽  
Conjugative Transfer ◽  
Rhizobium Etli ◽  
Wild Type ◽  
Specific System ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Symbiotic Plasmid ◽  
Recombination System ◽  
Attachment Sites

ABSTRACT Plasmid p42a from Rhizobium etli CFN42 is self-transmissible and indispensable for conjugative transfer of the symbiotic plasmid (pSym). Most pSym transconjugants also inherit p42a. pSym transconjugants that lack p42a always contain recombinant pSyms, which we designated RpSyms*. RpSyms* do not contain some pSym segments and instead have p42a sequences, including the replication and transfer regions. These novel recombinant plasmids are compatible with wild-type pSym, incompatible with p42a, and self-transmissible. The symbiotic features of derivatives simultaneously containing a wild-type pSym and an RpSym* were analyzed. Structural analysis of 10 RpSyms* showed that 7 shared one of the two pSym-p42a junctions. Sequencing of this common junction revealed a 53-bp region that was 90% identical in pSym and p42a, including a 5-bp central region flanked by 9- to 11-bp inverted repeats reminiscent of bacterial and phage attachment sites. A gene encoding an integrase-like protein (intA) was localized downstream of the attachment site on p42a. Mutation or the absence of intA abolished pSym transfer from a recA mutant donor. Complementation with the wild-type intA gene restored transfer of pSym. We propose that pSym-p42a cointegration is required for pSym transfer; cointegration may be achieved either through homologous recombination among large reiterated sequences or through IntA-mediated site-specific recombination between the attachment sites. Cointegrates formed through the site-specific system but resolved through RecA-dependent recombination or vice versa generate RpSyms*. A site-specific recombination system for plasmid cointegration is a novel feature of these large plasmids and implies that there is unique regulation which affects the distribution of pSym in nature due to the role of the cointegrate in conjugative transfer.

scholarly journals Identification of the Telomere elongation mutation in Drosophila

2018 ◽  
Author(s):  
Hemakumar M. Reddy ◽  
Thomas A. Randall ◽  
Radmila Capkova Frydrychova ◽  
James M. Mason
Keyword(s):  
Drosophila Melanogaster ◽  
Next Generation Sequencing ◽  
Ltr Retrotransposons ◽  
Wild Type ◽  
Dominant Mutation ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Telomere Elongation ◽  
Drosophila Genetic Reference Panel ◽  
Generation Sequencing

Background. Telomeres in Drosophila melanogaster are similar to those of other eukaryotes in terms of their function, although they are formed by non-LTR retrotransposons instead of telomerase-based short repeats. The length of the telomeres in Drosophila depends on the number of copies of these transposable elements. A dominant mutation, Tel1, causes a several-fold elongation of telomeres. Methods. In this study we identified the Tel1 mutation by a combination of transposon-induced, site-specific recombination and next generation sequencing. Results. Recombination located Tel1 to a 15 kb region in 92A. Comparison of the DNA sequence in this region with the Drosophila Genetic Reference Panel of wild type genomic sequences delimited Tel1 to a 3 bp deletion inside intron 8 of Ino80. Discussion. The mapped Tel1 mutation (3-bp deletion found in Ino80) did not appear to affect the quantity or length of the Ino80 transcript. Tel1 causes a significant reduction in transcripts of CG18493, a gene nested in an intron 8 of Ino80, which is expressed in ovaries and expected to encode a serine-type peptidase.

scholarly journals Role of attP in Integrase-Mediated Integration of the Shigella Resistance Locus Pathogenicity Island of Shigella flexneri

2004 ◽  
Vol 48 (3) ◽  
pp. 1028-1031 ◽  
Author(s):  
Sally A. Turner ◽  
Shelley N. Luck ◽  
Harry Sakellaris ◽  
Kumar Rajakumar ◽  
Ben Adler
Keyword(s):  
Shigella Flexneri ◽  
Pathogenicity Island ◽  
Resistance Locus ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Independent Site

ABSTRACT The Shigella resistance locus (SRL) pathogenicity island (PAI) in Shigella spp. mediates resistance to streptomycin, ampicillin, chloramphenicol, and tetracycline. It can be excised from the chromosome via site-specific recombination mediated by the P4-related int gene. Here, we show that SRL PAI attP is capable of RecA-independent, site-specific, int-mediated integration into two bacterial tRNA attB sites.

Characterization of Holliday structures in FLP protein-promoted site-specific recombination

1990 ◽  
Vol 10 (1) ◽  
pp. 235-242
Author(s):  
L Meyer-Leon ◽  
R B Inman ◽  
M M Cox
Keyword(s):  
Reaction Rate ◽  
Reaction Pathway ◽  
Reaction Intermediates ◽  
Wild Type ◽  
Site Specific ◽  
Site Specific Recombination ◽  
Isomerization Process ◽  
Previous Demonstration ◽  
Rate Limiting

Holliday structures are formed in the course of FLP protein-promoted site-specific recombination. Here, we demonstrate that Holliday structures are formed in reactions involving wild-type substrates and that they are kinetically competent with respect to the overall reaction rate. Together with a previous demonstration of chemical competence (L. Meyer-Leon, L.-C. Huang, S. W. Umlauf, M. M. Cox, and R. B. Inman, Mol. Cell. Biol. 8:3784-3796, 1988), Holliday structures therefore meet all criteria necessary to establish that they are obligate reaction intermediates in FLP-mediated site-specific recombination. In addition, kinetic evidence suggests that two distinct forms of the Holliday intermediate are present in the reaction pathway, interconverted in an isomerization process that is rate limiting at 0 degree C.

The role of himA and xis in lambda site-specific recombination

1980 ◽  
Vol 138 (3) ◽  
pp. 503-512 ◽  
Author(s):  
Susan Gottesman ◽  
Ken Abremski
Keyword(s):  
Site Specific ◽  
Site Specific Recombination

Site-specific Recombination of Bacteriophage  : The Role of Host Gene Products

1979 ◽  
Vol 43 (0) ◽  
pp. 1121-1126 ◽  
Author(s):  
H. I. Miller ◽  
A. Kikuchi ◽  
H. A. Nash ◽  
R. A. Weisberg ◽  
D. I. Friedman
Keyword(s):  
Host Gene ◽  
Gene Products ◽  
Site Specific ◽  
Site Specific Recombination

Role of Gin and FIS in Site-specific Recombination

1993 ◽  
Vol 58 (0) ◽  
pp. 505-513 ◽  
Author(s):  
P. Merker ◽  
G. Muskhelishvili ◽  
A. Deufel ◽  
K. Rusch ◽  
R. Kahmann
Keyword(s):  
Site Specific ◽  
Site Specific Recombination

Human acetyl-CoA:glucosamine-6-phosphate N-acetyltransferase 1 has a relaxed donor specificity and transfers acyl groups up to four carbons in length

2016 ◽  
Vol 94 (2) ◽  
pp. 197-204 ◽  
Author(s):  
Inka Brockhausen ◽  
Dileep G. Nair ◽  
Min Chen ◽  
Xiaojing Yang ◽  
John S. Allingham ◽  
...  
Keyword(s):  
Amino Acids ◽  
Articular Cartilage ◽  
Enzymatic Synthesis ◽  
Biomedical Applications ◽  
Binding Pocket ◽  
Wild Type ◽  
Acetyl Coenzyme A ◽  
Site Specific ◽  
Acetyl Group

Glucosamine-6-phosphate N-acetyltransferase1 (GNA1) catalyses the transfer of an acetyl group from acetyl coenzyme A (AcCoA) to glucosamine-6-phosphate (GlcN6P) to form N-acetylglucosamine-6-phosphate (GlcNAc6P), which is an essential intermediate in UDP-GlcNAc biosynthesis. An analog of GlcNAc, N-butyrylglucosamine (GlcNBu) has shown healing properties for bone and articular cartilage in animal models of arthritis. The goal of this work was to examine whether GNA1 has the ability to transfer a butyryl group from butyryl-CoA to GlcN6P to form GlcNBu6P, which can then be converted to GlcNBu. We developed fluorescent and radioactive assays and examined the donor specificity of human GNA1. Acetyl, propionyl, n-butyryl, and isobutyryl groups were all transferred to GlcN6P, but isovaleryl-CoA and decanoyl-CoA did not serve as donor substrates. Site-specific mutants were produced to examine the role of amino acids potentially affecting the size and properties of the AcCoA binding pocket. All of the wild type and mutant enzymes showed activities of both acetyl and butyryl transfer and can therefore be used for the enzymatic synthesis of GlcNBu for biomedical applications.

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