Microtubule Affinity Regulating Kinase Activity in Living Neurons Was Examined by a Genetically Encoded Fluorescence Resonance Energy Transfer/Fluorescence Lifetime Imaging-based Biosensor

Timm, Thomas; Kries, Jens Peter von; Li, Xiaoyü; Zempel, Hans; Mandelkow, Eckhard; Mandelkow, E.

doi:10.1074/jbc.m111.257865

Cited by 42 publications

(40 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A second compound with higher IC 50 for DYRK1B but with no specificity for CLK kinases NCGC00185984 was not as active in blocking NKX3.1 degradation. A MARK2 inhibitor, 39621, was used because MARK2 was one of the kinases identified in the second round kinome library screening (24). Compound 39621 was substantially less active as an inhibitor of NKX3.1 degradation than the DYRK1B inhibitors.…”

Section: Resultsmentioning

confidence: 99%

The Tumor Suppressor NKX3.1 Is Targeted for Degradation by DYRK1B Kinase

Song

Silva²,

Koller

et al. 2015

Molecular Cancer Research

View full text Add to dashboard Cite

NKX3.1 is a prostate specific homeodomain protein and tumor suppressor whose expression is reduced in the earliest phases of prostatic neoplasia. NKX3.1 expression is not only diminished by genetic loss and methylation, but the protein itself is a target for accelerated degradation caused by inflammation that is common in the aging prostate gland. NKX3.1 degradation is activated by phosphorylation at C-terminal serine residues that mediate ubiquitination and protein turnover. Since NKX3.1 is haploinsufficient, strategies to increase its protein stability could lead to new therapies. Here, a high throughput screen was developed using a siRNA library for kinases that mediate NKX3.1 degradation. This approach identified several candidates of which DYRK1B, a kinase that is subject to gene amplification and overexpression in other cancers, had the greatest impact on NKX3.1 half-life. Mechanistically, NKX3.1 and DYRK1B were shown to interact via the DYRK1B kinase domain. In addition, an in vitro kinase assay showed that DYRK1B phosphorylated NKX3.1 at serine185, a residue critical for NKX3.1 steady state turnover. Lastly, small molecule inhibitors of DYRK1B prolonged NKX3.1 half-life. Thus, DYRK1B is a target for enzymatic inhibition in order to increase cellular NKX3.1. Implications DYRK1B is a promising and novel kinase target for prostate cancer treatment mediated by enhancing NKX3.1 levels.

show abstract

Section: Resultsmentioning

confidence: 99%

The Tumor Suppressor NKX3.1 Is Targeted for Degradation by DYRK1B Kinase

Song

Silva²,

Koller

et al. 2015

Molecular Cancer Research

View full text Add to dashboard Cite

show abstract

“…2.1a); however, the upstream events activating these signaling cascades have not been clarified (Timm et al 2003(Timm et al , 2008a. The protein structure of the kinase domain with the UBA domain has been determined for MARK1, MARK2, and MARK3 Marx et al 2006;Murphy et al 2007), and a candidate for a PAR-1-specific inhibitor has also been reported (Timm et al 2011). It is also noteworthy that CagA, the toxin of Helicobacter pylori, has been shown to inhibit PAR-1 kinase activity by specifically binding to the substrate-binding pocket of the kinase domain (Nesic et al 2010).…”

Section: Essential Features Of Par-1mentioning

confidence: 96%

PAR-1 Kinase and Cell Polarity

Suzuki

2015

Cell Polarity 1

View full text Add to dashboard Cite

PAR-1 kinase was identified as one of the protein products of the par (partition defective) genes essential for the establishment of the anterior-posterior polarity of the Caenorhabditis elegans one-cell embryo. Subsequent studies have revealed the ubiquitous importance of the kinase in regulating cell polarity observed in various biological contexts of many organisms. As a membranelocalized kinase, PAR-1 antagonizes the atypical protein kinase C (aPKC)/PAR-3/PAR-6 complex to establish the mutually exclusive membrane domains along the polarity axis. Based on this asymmetric membrane localization, its kinase activity is utilized to phosphorylate various other target proteins that regulate cellular functions. The major function of PAR-1 is the regulation of microtubule dynamics, of which one of the underlying mechanisms was identified as the microtubule affinityregulating kinase (MARK) activity in mammalian neurons. In addition, accumulating data have revealed the presence of various kinds of other target proteins involved in the regulation of actin cytoskeleton and protein stability. Although the significant versatility of PAR-1 has made it increasingly difficult to obtain a simple view of its function, the evolutional insight on PAR-1 provides a powerful tool for integrating increasing data on this kinase in the light of cell polarity.

show abstract

“…Further enhancement of the dynamic range could facilitate this important application in cellular context to complement FRET-based inhibitor screens using synthetic sensors outside of the cellular context (e.g., Gratz et al, 2011). Another application of the genetically encoded sensors where we expect to see further advancement is inhibitor candidate validation (e.g., Timm et al, 2011), including validation of inhibitors’ isoform specificity (Tsalkova et al, 2012), and analysis of pharmacokinetics in in vitro or in vivo models (Nobis et al, 2013).…”

Section: Discussionmentioning

confidence: 99%

Genetically Encoded Fluorescent Biosensors for Live-Cell Visualization of Protein Phosphorylation

Oldach

Zhang

2014

Chemistry & Biology

View full text Add to dashboard Cite

Fluorescence-based, genetically encodable biosensors are widely used tools for real-time analysis of different biological process. Over the last few decades the number of available genetically encodable biosensors and the types of processes they can monitor has increased rapidly. In this review we aim to introduce the reader to general principles and best practices in biosensor development and highlight some of the ways in which biosensors can be used to illuminate outstanding questions of biological function. Specifically, we will focus on sensors developed for monitoring kinase activity and use them to illustrate some common considerations for biosensor design. We will describe several uses to which kinase and second-messenger biosensors have been put, and conclude with considerations for the use of biosensors once they are developed. Overall, as fluorescence-based biosensors continue to diversify and improve we expect them to continue to be widely used as reliable and fruitful tools for gaining deeper insights into cellular and organismal function.

show abstract

Microtubule Affinity Regulating Kinase Activity in Living Neurons Was Examined by a Genetically Encoded Fluorescence Resonance Energy Transfer/Fluorescence Lifetime Imaging-based Biosensor

Cited by 42 publications

References 43 publications

The Tumor Suppressor NKX3.1 Is Targeted for Degradation by DYRK1B Kinase

The Tumor Suppressor NKX3.1 Is Targeted for Degradation by DYRK1B Kinase

PAR-1 Kinase and Cell Polarity

Genetically Encoded Fluorescent Biosensors for Live-Cell Visualization of Protein Phosphorylation

Contact Info

Product

Resources

About