LOV-based reporters for fluorescence imaging

Buckley, Anthony M.; Petersen, J.; Roe, Andrew J.; Douce, Gill; Christie, John M.

doi:10.1016/j.cbpa.2015.05.011

Cited by 106 publications

(109 citation statements)

References 57 publications

Supporting

Mentioning

102

Contrasting

Order By: Relevance

“…For instance, a recent publication has demonstrated that the subset of F-actin that is recognized by diverse ABDs can be altered by the fluorescent protein or the linker sequence between the ABD and the fluorescent protein (Lemieux et al, 2014). New applications for fluorescent labeling of proteins have become available recently, such as the use of Y-FAST, a small monomeric protein tag enabling reversible binding and activation of a cell-permeant and nontoxic fluorogenic molecule (Plamont et al, 2016), or the flavoprotein improved LOV (iLOV), a fluorescent protein based on the light, oxygen or voltage domain (LOV) from a variety of sources (Buckley et al, 2015). Both molecules offer advantages over GFP and other related fluorescent reporters owing to their relatively small size.…”

Section: Discussionmentioning

confidence: 99%

Actin visualization at a glance

Melak

Plessner

Grosse

2017

Journal of Cell Science

198

200

View full text Add to dashboard Cite

There was an error published in J. Cell Sci. 130,[525][526][527][528][529][530] Unfortunately, the diagram of the actin filament in the poster was accidentally reflected during the creation of the figure. As a result, the filament was illustrated as a left-handed rather than a right-handed F-actin helix in the original version of this article. The online version of the article has been corrected accordingly. This change has no impact on the concepts presented in this article.The authors apologise to the readers for any confusion that this error might have caused. 1688 Journal of Cell Science CELL SCIENCE AT A GLANCE Actin visualization at a glanceMichael Melak, Matthias Plessner and Robert Grosse* ABSTRACT Actin functions in a multitude of cellular processes owing to its ability to polymerize into filaments, which can be further organized into higher-order structures by an array of actin-binding and regulatory proteins. Therefore, research on actin and actin-related functions relies on the visualization of actin structures without interfering with the cycles of actin polymerization and depolymerization that underlie cellular actin dynamics. In this Cell Science at a Glance and the accompanying poster, we briefly evaluate the different techniques and approaches currently applied to analyze and visualize cellular actin structures, including in the nuclear compartment. Referring to the gold standard F-actin marker phalloidin to stain actin in fixed samples and tissues, we highlight methods for visualization of actin in living cells, which mostly apply the principle of genetically fusing fluorescent proteins to different actin-binding domains, such as LifeAct, utrophin and F-tractin, as well as antiactin-nanobody technology. In addition, the compound SiR-actin and the expression of GFP-actin are also applicable for various types of live-cell analyses. Overall, the visualization of actin within a physiological context requires a careful choice of method, as well as a tight control of the amount or the expression level of a given detection probe in order to minimize its influence on endogenous actin dynamics.

show abstract

Section: Discussionmentioning

confidence: 99%

Actin visualization at a glance

Melak

Plessner

Grosse

2017

Journal of Cell Science

198

200

View full text Add to dashboard Cite

show abstract

“…The key advantages of using phiLOV2.1 as a reporter are its stability over a wide range of pH values, its molecular oxygen independency and its small size (12.1 kDa). Conversely, photobleaching, and weaker signals are disadvantages (Buckley et al, 2015). Recently, protein translocation into mammalian cells through the type 3 secretion system of Shigella flexneri was successfully visualized using phiLOV2.1 (Gawthorne et al, 2016).…”

Section: Discussionmentioning

confidence: 99%

“…Recently, protein translocation into mammalian cells through the type 3 secretion system of Shigella flexneri was visualized using phiLOV2.1, an improved version of LOV (Gawthorne et al, 2016). The LOV domain is responsible for the fluorescence properties of the plant blue-light receptor kinases called phototropins, regulated either by Light, Oxygen or Voltage (Huala et al, 1997;Buckley et al, 2015). Compared with LOV phiLOV2.1 has increased fluorescence and photostability (Christie et al, 2012a,b).…”

Section: Introductionmentioning

confidence: 99%

Application of phiLOV2.1 as a fluorescent marker for visualization of Agrobacterium effector protein translocation

et al. 2018

View full text Add to dashboard Cite

Agrobacterium tumefaciens can genetically transform plants by translocating a piece of oncogenic DNA, called T-DNA, into host cells. Transfer is mediated by a type IV secretion system (T4SS). Besides the T-DNA, which is transferred in a single-stranded form and at its 5' end covalently bound to VirD2, several other effector proteins (VirE2, VirE3, VirD5, and VirF) are translocated into the host cells. The fate and function of the translocated proteins inside the host cell are only partly known. Therefore, several studies were conducted to visualize the translocation of the VirE2 protein. As GFP-tagged effector proteins are unable to pass the T4SS, other approaches like the split GFP system were used, but these require specific transgenic recipient cells expressing the complementary part of GFP. Here, we investigated whether use can be made of the photostable variant of LOV, phiLOV2.1, to visualize effector protein translocation from Agrobacterium to non-transgenic yeast and plant cells. We were able to visualize the translocation of all five effector proteins, both to yeast cells, and to cells in Nicotiana tabacum leaves and Arabidopsis thaliana roots. Clear signals were obtained that are easily distinguishable from the background, even in cases in which by comparison the split GFP system did not generate a signal.

show abstract

“…All systems that rely on endogenous chromophores are based on one of three compounds: flavin mononucleotide (FMN), biliverdin, or bilirubin (Figures and , Table ). Systems that rely on complexation of FMN are typically non‐covalent complexes that fluoresce in the cyan‐green wavelength range . These systems are based on bacterial photoreceptors comprised of light, oxygen, and voltage (LOV) sensing domains, which form a reversible covalent bond with FMN via a conserved cysteine residue over the course of their photocycle.…”

Section: Introductionmentioning

confidence: 99%

Fluorogenic Protein‐Based Strategies for Detection, Actuation, and Sensing

Gautier

Tebo

2018

BioEssays

View full text Add to dashboard Cite

Fluorescence imaging has become an indispensable tool in cell and molecular biology. GFP-like fluorescent proteins have revolutionized fluorescence microscopy, giving experimenters exquisite control over the localization and specificity of tagged constructs. However, these systems present certain drawbacks and as such, alternative systems based on a fluorogenic interaction between a chromophore and a protein have been developed. While these systems are initially designed as fluorescent labels, they also present new opportunities for the development of novel labeling and detection strategies. This review focuses on new labeling protocols, actuation methods, and biosensors based on fluorogenic protein systems.

show abstract

LOV-based reporters for fluorescence imaging

Cited by 106 publications

References 57 publications

Actin visualization at a glance

Actin visualization at a glance

Application of phiLOV2.1 as a fluorescent marker for visualization of Agrobacterium effector protein translocation

Fluorogenic Protein‐Based Strategies for Detection, Actuation, and Sensing

Contact Info

Product

Resources

About