Minimotif Miner: A Computational Tool to Investigate Protein Function, Disease, and Genetic Diversity

Schiller, Martin

doi:10.1002/0471140864.ps0212s48

Cited by 8 publications

(6 citation statements)

References 30 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Interestingly, the characteristic GSR repeats within this sequence have 6 serines that match consensus GSK3β phosphorylation sites (phosphorylation cluster-1, P1). While in-silico analysis with different phosphorylation prediction algorithms (Obenauer et al, 2003; Schiller, 2007) reveal additional potential GSK3β sites (phosphorylation cluster 2, P2, Fig. 1F), their incompatible sequence context precluded efficient peptide retrieval for MS/MS analysis.…”

Section: Resultsmentioning

confidence: 99%

Skin Stem Cells Orchestrate Directional Migration by Regulating Microtubule-ACF7 Connections through GSK3β

Shen

Oristian

et al. 2011

Cell

168

230

View full text Add to dashboard Cite

SUMMARY Homeostasis and wound-healing rely on stem cells (SCs) whose activity and directed migration are often governed by Wnt signaling. In dissecting how this pathway integrates with the necessary downstream cytoskeletal dynamics, we discovered that GSK3β directly phosphorylates ACF7, a >500kd microtubule-actin crosslinking protein abundant in hair follicle stem cells (HF-SCs). We map ACF7’s GSK3β sites to the microtubule-binding domain and show that phosphorylation uncouples ACF7 from microtubules. Phosphorylation-refractile ACF7 rescues overall microtubule architecture, but phosphorylation-constitutive mutants do not. Neither mutant rescues polarized movement, revealing that phospho-regulation must be dynamic. This circuitry is physiologically relevant, depending upon polarized GSK3β inhibition at the migrating front of SCs/progeny streaming from HFs during wound-repair. Moreover, only ACF7 and not GSKβ-refractile-ACF7 restore polarized microtubule-growth and SC-migration to ACF7-null skin. Our findings provide insights into how this conserved spectraplakin integrates signaling, cytoskeletal dynamics and polarized locomotion of somatic SCs.

show abstract

Section: Resultsmentioning

confidence: 99%

Skin Stem Cells Orchestrate Directional Migration by Regulating Microtubule-ACF7 Connections through GSK3β

Shen

Oristian

et al. 2011

Cell

168

230

View full text Add to dashboard Cite

show abstract

“…There is much still to learn. An analysis of the P2X7R sequence using Minimotif Miner Scan [85][86][87] and focusing on the C terminus has allowed us to find many new and previously undiscovered motifs and domains listed in Table 1.…”

Section: What Is Expected For the Future?mentioning

confidence: 99%

C terminus of the P2X7 receptor: treasure hunting

Costa-Junior

Vieira

Coutinho‐Silva

2011

Purinergic Signalling

107

View full text Add to dashboard Cite

P2X receptor (P2XR) is a family of the ATPgated ion channel family and can permeabilize the plasma membrane to small cations such as potassium, sodium, and calcium, resulting in cellular depolarization. There are seven P2XR that have been described and cloned, with 45% identity in amino acid sequence. Each P2X receptors has two transmembrane domains that are separated by an extracellular loop and an intracellular N and C terminus. Unlike the other P2X receptors, the P2X7R has a larger C terminus with an extra 200 amino acid residues compared with the other receptors. The C terminus of the P2X7R has been implicated in regulating receptor function including signaling pathway activation, cellular localization, proteinprotein interactions, and post-translational modification (PTM). In the present review, we discuss the role of the P2X7R C terminus in regards to receptor function, describe the specific domains and motifs found therein and compare the C terminus sequence with others proteins to discover predicted domains or sites of PTM.

show abstract

“…We previously used a web-based application, Minimotif Miner (MnM 1.0), to search within EFF-1’s cytoplasmic domain for instances of short peptide motifs (<15 residues) of known function [ 51 – 53 ]. This analysis revealed two candidate 14-3-3-binding motifs as the highest scoring potentially functional sites.…”

Section: Introductionmentioning

confidence: 99%

The EFF-1A Cytoplasmic Domain Influences Hypodermal Cell Fusions in C. elegans But Is Not Dependent on 14-3-3 Proteins

et al. 2016

View full text Add to dashboard Cite

BackgroundRegulatory and biophysical mechanisms of cell-cell fusion are largely unknown despite the fundamental requirement for fused cells in eukaryotic development. Only two cellular fusogens that are not of clear recent viral origin have been identified to date, both in nematodes. One of these, EFF-1, is necessary for most cell fusions in Caenorhabditis elegans. Unregulated EFF-1 expression causes lethality due to ectopic fusion between cells not developmentally programmed to fuse, highlighting the necessity of tight fusogen regulation for proper development. Identifying factors that regulate EFF-1 and its paralog AFF-1 could lead to discovery of molecular mechanisms that control cell fusion upstream of the action of a membrane fusogen. Bioinformatic analysis of the EFF-1A isoform’s predicted cytoplasmic domain (endodomain) previously revealed two motifs that have high probabilities of interacting with 14-3-3 proteins when phosphorylated. Mutation of predicted phosphorylation sites within these motifs caused measurable loss of eff-1 gene function in cell fusion in vivo. Moreover, a human 14-3-3 isoform bound to EFF-1::GFP in vitro. We hypothesized that the two 14-3-3 proteins in C. elegans, PAR-5 and FTT-2, may regulate either localization or fusion-inducing activity of EFF-1.Methodology/Principal FindingsTiming of fusion events was slightly but significantly delayed in animals unable to produce full-length EFF-1A. Yet, mutagenesis and live imaging showed that phosphoserines in putative 14-3-3 binding sites are not essential for EFF-1::GFP accumulation at the membrane contact between fusion partner cells. Moreover, although the EFF-1A endodomain was required for normal rates of eff-1-dependent epidermal cell fusions, reduced levels of FTT-2 and PAR-5 did not visibly affect the function of wild-type EFF-1 in the hypodermis.Conclusions/SignificanceDeletion of the EFF-1A endodomain noticeably affects the timing of hypodermal cell fusions in vivo. However, prohibiting phosphorylation of candidate 14-3-3-binding sites does not impact localization of the fusogen. Hypodermal membrane fusion activity persists when 14-3-3 expression levels are reduced.

show abstract

Minimotif Miner: A Computational Tool to Investigate Protein Function, Disease, and Genetic Diversity

Cited by 8 publications

References 30 publications

Skin Stem Cells Orchestrate Directional Migration by Regulating Microtubule-ACF7 Connections through GSK3β

Skin Stem Cells Orchestrate Directional Migration by Regulating Microtubule-ACF7 Connections through GSK3β

C terminus of the P2X7 receptor: treasure hunting

The EFF-1A Cytoplasmic Domain Influences Hypodermal Cell Fusions in C. elegans But Is Not Dependent on 14-3-3 Proteins

Contact Info

Product

Resources

About