IS911-mediated transpositional recombination in vitro

Journal of Molecular Biology
P PolardM Chandler

Abstract

A cell-free system is described that accomplishes an unusual type of transposition/recombination involving the bacterial insertion sequence IS911. Using a plasmid substrate carrying a derivative of IS911, we show that bacterial cell extracts enriched for the IS911 transposase, OrfAB, carry out a single-strand cleavage and transfer reaction. This results in the formation of a figure-eight molecule in which a single strand of the element is circularized, faithfully reproducing an event previously detected in vivo. Moreover, when presented with a figure-eight substrate, OrfAB is capable of "reversing" strand transfer. This activity is equivalent to the "disintegration" reaction carried out by retroviral integrases. We demonstrate that the domain of OrfAB responsible for this catalytic activity is located in the carboxy-terminal region of the protein, since a peptide composed of this region retains disintegration activity. The OrfAB-mediated excision-circularization process previously observed in vivo was proposed to proceed via a figure-eight intermediate by circularization of the second transposon strand. The absence of transposon circles in cell-free reaction suggests either that the figure-eight form is not an intermediate or t...Continue Reading

Citations

May 13, 1999·Molecular Microbiology·B Ton-HoangM Chandler
Mar 28, 1998·The EMBO Journal·B Ton-HoangM Chandler
Apr 1, 2009·Antimicrobial Agents and Chemotherapy·Laurent PoirelPatrice Nordmann
Jul 1, 2008·Journal of Bacteriology·P RousseauM Chandler
Jul 30, 2004·The EMBO Journal·T K AuRasika M Harshey
Nov 3, 2006·Molecular Microbiology·Erwan GueguenMick Chandler
Nov 18, 2003·Annual Review of Genetics·Finbarr Hayes
Oct 23, 2012·Mobile Genetic Elements·Jean Challacombe, Cheryl Kuske
Nov 5, 1999·Annual Review of Microbiology·L HarenM Chandler
Nov 21, 2007·Journal of Molecular Biology·Mónika SzabóFerenc Olasz
Feb 6, 2004·Molecular Microbiology·Celine LootMichael Chandler
Aug 13, 2004·Molecular Microbiology·Catherine TurlanMichael Chandler
Mar 21, 2007·Molecular Microbiology·Philippe RousseauMick Chandler
Oct 5, 2001·The Journal of Biological Chemistry·S MarsinP Polard
Jan 17, 2017·Mobile Genetic Elements·Vandan Shah, Jin Ryoun Kim
Jun 27, 1998·Molecular Microbiology·F OlaszW Arber
Oct 30, 2019·Nature Structural & Molecular Biology·George E GhanimDonald C Rio
Apr 12, 2000·Molecular Microbiology·C TurlanM Chandler
Jun 25, 2015·Microbiology Spectrum·Rasika M Harshey
Sep 8, 1998·Microbiology and Molecular Biology Reviews : MMBR·J Mahillon, M Chandler
Nov 9, 2007·Journal of Molecular Biology·Kathryn M LembergTania A Baker

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.