Dynamic, Structural, and Regulatory Aspects of Λ Site-Specific Recombination

Landy, Arthur

doi:10.1146/annurev.bi.58.070189.004405

Cited by 565 publications

(287 citation statements)

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“…A collection of yeast strains that contain plasmid pairs expressing fusion proteins with a spectrum of interaction strengths [pPC97 (GAL4-DB, LEU2), pPC97-CYH2 s and pPC86 (GAL4-AD, TRP1)] were used as controls (53,(56)(57)(58)(59). The control plasmids pDBIeu and pEXP-AD507 contain only the Gal4 DNA-binding domain (BD) and the Gal4 activating-domain (AD) respectively.…”

Section: Yeast Strains For Two-hybrid Assaymentioning

confidence: 99%

A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

et al. 2007

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Although plasma membrane domains such as caveolae provide an organizing principle for signaling pathways and cholesterol homeostasis in the cell, relatively little is known regarding specific mechanisms whereby intracellular lipid binding proteins are targeted to caveolae. Therefore, the interaction between caveolin-1 and sterol carrier protein-2 (SCP-2), a protein that binds and transfers both cholesterol and signaling lipids (e.g. phosphatidylinositides, sphingolipids), was examined by yeast two-hybrid, in vitro binding, and fluorescence resonance energy transfer analyses (FRET). Results of the in vivo and in vitro assays identified for the first time the N-terminal aa1-32 amphipathic α-helix of SCP-2 functionally interacted with caveolin-1. This interaction was independent of the classic caveolin-1 scaffolding domain in which many signaling proteins interact. Instead, SCP-2 bound caveolin-1 through a new domain identified in the N-terminal domain of caveolin-1 between amino acids 32-55. Modeling studies suggested that electrostatic interactions between the SCP-2 N-terminal aa1-32 amphipathic α-helical domain (cationic, positively charged face) and the caveolin-1 N-terminal aa33-59 α-helix (anionic, negatively charged face) may significantly contribute to this interaction. These findings provide new insights on how SCP-2 enhances cholesterol retention within the cell as well as regulates the distribution of signaling lipids such as phosphoinositides and sphingolipids at plasma membrane caveolae.Keywords sterol carrier protein-2; caveolae; caveolin-1; caveolae; cholesterol; signaling Increasing evidence indicates that cholesterol found at the cell surface plasma membrane (PM) is not randomly distributed, but instead organized into both transbilayer (1,2) and lateral (3,4) cholesterol-rich (and/or sphingolipid-rich) domains that adopt a unique, liquidordered structural organization (4-7). It has been postulated that this self-assembling property of cholesterol (and also sphingolipids) into domains in turn forms the structural basis for selective membrane protein organization (8). Support for this hypothesis is from numerous studies demonstrating that many PM proteins are functionally organized into lipid rafts and/or caveolae, a sub-fraction of lipid rafts that have proven to be a remarkably stable † Acknowledgments: This work was supported in part by the USPHS National Institutes of Health GM31651 (FS and AK) and GM62326 (JMB).

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Section: Yeast Strains For Two-hybrid Assaymentioning

confidence: 99%

A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

et al. 2007

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“…While for some reactions, the free energy of DNA supercoiling favours the products, stabilization of the product-containing complex probably contributes significantly to the directionality of many reactions. For example, in l site-specific recombination, the reaction direction is controlled by involving two overlapping sets of proteins under different physiological conditions, preferentially stabilizing one product set over another (Landy 1989). Another example is the strand transfer step of transposition reactions which should reach equilibrium at near 50%, at best, without the product stabilization that apparently pushes the reaction toward completion (Surette et al 1987;Haniford et al 1991).…”

Section: Energetic Considerations and Recombination Complexesmentioning

confidence: 99%

“…A number of recent reviews have addressed a variety of mechanistic aspects of these reactions (Berg & Howe 1989;Landy 1989;Mizuuchi 1992a;Stark et al 1992). In this review, I will consider several aspects of the chemical steps of polynucleotidyl transfer by these recombination enzymes.…”

Section: Introductionmentioning

confidence: 99%

Polynucleotidyl transfer reactions in site‐specific DNA recombination

1997

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Site-specific DNA rearrangement reactions are widespread among organisms. They are used, for example, by vertebrates to boost immune response diversity, and in turn by parasitic organisms to evade the host immune system by surface antigen switching. Parasitic genetic elements ubiquitous to most organisms invade new host genomic sites by a variety of types of site-specific recombination. Polynucleotidyl transfer reactions are central to these DNA recombination reactions. The recombinase of each reaction system that 'catalyses' such chemical reactions at specific DNA sites are apparently designed to accomplish unique DNA geometrical specificity, or delicate control over the extent or direction of the reaction, with the sacrifice of protein turnover. Here we discuss our current understanding of several issues that relate to the polynucleotidyl transfer steps in several of the better studied site-specific recombination reactions.

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“…This rapid, directional, and highly efficient cloning system is based on site-specific recombination mediated by the recombination machinery of bacteriophage lambda (Landy, 1989) and has been adapted for in vitro use in standard molecular biology protocols (Hartley et al, 2000;Walhout et al, 2000).…”

Section: Overview Of the Gateway System Of Recombination-based Cloningmentioning

confidence: 99%

A suite of Gateway^® cloning vectors for high‐throughput genetic analysis in Saccharomyces cerevisiae

Alberti

Gitler

Lindquist

2007

Yeast

427

419

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In the post-genomic era, academic and biotechnological research is increasingly shifting its attention from single proteins to the analysis of complex protein networks. This change in experimental design requires the use of simple and experimentally tractable organisms, such as the unicellular eukaryote Saccharomyces cerevisiae, and a range of new high-throughput techniques. The Gateway  system has emerged as a powerful high-throughput cloning method that allows for the in vitro recombination of DNA with high speed, accuracy and reliability. Two Gateway-based libraries of overexpression plasmids containing the entire complement of yeast open reading frames (ORFs) have recently been completed. In order to make use of these powerful resources, we adapted the widely used pRS series of yeast shuttle vectors for use in Gateway-based cloning. The resulting suite of 288 yeast Gateway vectors is based upon the two commonly used GPD and GAL1 promoter expression systems that enable expression of ORFs, either constitutively or under galactoseinducible conditions. In addition, proteins of interest can be fused to a choice of frequently used N-or C-terminal tags, such as EGFP, ECFP, EYFP, Cerulean, monomeric DsRed, HA or TAP. We have made this yeast Gateway  vector kit available to the research community via the non-profit Addgene Plasmid Repository (http://www.addgene.org/yeast gateway).

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Dynamic, Structural, and Regulatory Aspects of Λ Site-Specific Recombination

Cited by 565 publications

References 0 publications

A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

Polynucleotidyl transfer reactions in site‐specific DNA recombination

A suite of Gateway^® cloning vectors for high‐throughput genetic analysis in Saccharomyces cerevisiae

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Dynamic, Structural, and Regulatory Aspects of Λ Site-Specific Recombination

Cited by 565 publications

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A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

A New N-Terminal Recognition Domain in Caveolin-1 Interacts with Sterol Carrier Protein-2 (SCP-2)

Polynucleotidyl transfer reactions in site‐specific DNA recombination

A suite of Gateway® cloning vectors for high‐throughput genetic analysis in Saccharomyces cerevisiae

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Product

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A suite of Gateway^® cloning vectors for high‐throughput genetic analysis in Saccharomyces cerevisiae