Global ETD Search
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Showing 1 to 20 of 27 for “"Phosphomimetic"”.
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Characterization of a Phosphomimetic Mutant of the ALS Associated Protein TDP-43
… propensity. We expressed a recombinant S48E phosphomimetic mutation of TDP-43 with an N-terminal GFP fluorescent tag in <em>Escherichia coli</em> and induced aggregation by agitation. We examined the extent to which different DNA/RNAs and differing stoichiometric concentrations of these …
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Structural and Biochemical Investigation of Fanconi Anemia Pathway Activation
… mechanism of this process by employing a triple-phosphomimetic FANCI. First, I confirmed the stimulatory effect of phosphomimetic FANCI on D2-I ubiquitination through in vitro ubiquitination assays using purified proteins. To elucidate the structural basis of this stimulation, I visualized …
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Role of Phosphorylated Dicer1 In Tumor Progression
… of a genetically engineered mouse model of phosphomimetic <em>Dicer1</em> when combined with heterozygous <em>Kras</em> oncogenic background contributes to lung tumor progression <em>in vivo</em>. Mechanisms through which phosphomimetic <em>Dicer1</em> regulates tumor progression remain …
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Structural and Functional Effect of Phosphorylation On Ampa Receptors
… associated with activation of the receptor. The phosphomimetic receptor, on the other hand, shows no change in FRET associated with activation of the receptor. However, addition of the lipid sphingosine restores this change in FRET due to activation for the phosphomimetic receptor. Using single …
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Phosphorylation of The Smooth Muscle Master Splicing Regulator, RBPMS-A, Regulates its Splicing Activity
… activity. I, therefore, created a series of phosphomimetic (T/E) and non-phosphomimetic (T/A) mutants to explore what effect phosphorylation of these residues might have on splicing. Transfection of these mutant constructs showed that Thr113 and Thr118 are critical in regulating the splicing …
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A Novel Phosphorylation Site in the Telomeric Protein TRF2 is Regulated by the ATR Kinase and Plays a Role in Relieving Replication Stress at the Telomere
… In mouse cells, expression of TRF2 containing a phosphomimetic mutation at position 366 (S366E) caused dramatic telomere lengthening; in human cells, overexpression of S368 mutants did not reproduce the rapid telomere shortening caused by overexpression of wildtype TRF2, indicating that these …
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Regulatory mechanisms of MYO6 motor domain activity and tail interactions
… the cell surface, which upon expression of the phosphomimetic significantly increases in number. Co-expression of the phosphomimetic and wildtype MYO6 causes a re-distribution of their localization to the filopodia tip and stalk, respectively. I also identified the kinase that phosphorylates …
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Interplay Between Keratin and Vimentin Expression in Oral Cancer
… with plasmids encoding wild-type, PKCε-phosphomimetic, or unphosphorylatable versions of vimentin. Expression of vimentin was confirmed by western blot and immunofluorescence. Effects on cell growth and motility were determined using MTT, cell proliferation, and wound-closure assays. …
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Studies of Huntington's disease associated motor domain phosphorylation of kinesin-1
… in the processivity or ATPase activity of a phosphomimetic S175D construct or the non-phosphorylatable S175A construct. However, we did find that addition of a negative charge at S175 through phosphorylation or mutation led to a decreased stall force. Furthermore, polystyrene bead cargos …
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Regulation of SNARE-dependent fusion in an in vitro system
… unable to bind syntaxin 4 in vitro. Furthermore, phosphomimetic mutations were also introduced on both proteins and pull-down assays indicated that phosphorylated Munc18c is unable to interact with both syntaxin 4 and VAMP2, whereas phosphomimetic mutations in syntaxin 4 did not affect the …
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Regulation of Asymmetric Cell Divisions in the Developing Epidermis
… localized predominately to the cortex while the phosphomimetic protein localized strongly to spindle poles. In agreement with these data, use of a CDK-1 inhibitor was able to enhance the cortical localization of NuMA. Unexpectedly, the non-phosphorylatable form of NuMA did not require LGN to …
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Régulation du récepteur aux opioïdes de type delta par les protéines Homer1
… assays. We then generated truncated, alanine and phosphomimetic GSThDOP-CT mutants to investigate a potential Homer1 consensus binding motif. Whereas amino acids 339 to 354 of hDOP-CT were important to Homer1 binding, individual residues within this sequence were not identified as essential for …
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The Role of Traf6 Phosphorylation In Src/Traf6-Mediated Ikk, Jnk, Akt Activation and Tumorigenesis
… Akt, and JNK activation, which can be rescued by phosphomimetic TRAF6, but not by phosphorylation-dead TRAF6 mutant. Mechanistically, we show that TRAF6 phosphorylation is imperative for TRAF6 dimerization/oligomerization and its binding to E2 enzyme, UBC13. Significantly, we find that TRAF6 …
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Maintenance of Genomic Integrity by RSF1 Chromatin Remodeler
… from chromosome-alignment defects are rescued by phosphomimetic RSF1 mutants, but not by non-phosphorylatable mutants 31 IV. DISCUSSION 35 PART2. 43 I. INTRODUCTION 44 II. MATERIALS AND METHODS 46 K. Immunoprecipitation and in vitro binding assay 46 L. Cell fractionation 46 M. Chromosome spreading …
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Investigating galactose metabolism in Streptococcus pneumoniae
… non-phosphorylatable residue alanine (A) or the phosphomimetic aspartic acid (D) or glutamic acid (E). While the use of phosphomimetic residues proved somewhat challenging, the use of non-phosphorylatable alanine residues revealed that each of the three putative phosphorylation sites are required …
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Structural basis of membrane targeting and regulation of the innate immunity adaptor TIRAP by its phosphoinositide-binding motif
… PIPs to stabilize TIRAP's membrane interaction. Phosphomimetic mutation of Thr28 to Asp (T28D) as well as phosphorylation in Thr28 inhibit TIRAP PBM's binding to phosphoinositides by distorting the central helical structure of the peptide. More importantly, TIRAP T28D disrupt its subcellular …
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Characterization of cellular stress systems using biological mass spectrometry
… as a cell survival factor. With the help of phosphomimetic and non-phosphorylable mutants, we were able to decipher their effect on localization and progression through the cell cycle. Collectively, these studies manifest the power of MS-generated data to influence and guide many different …
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Camkii-Mediated Phosphorylation of the Inositol 1,4,5-Trisphosphate Receptor at Serine-150 Results in Decreased Channel Activity
… showed no decrease in activity. Conversely, the phosphomimetic channel displayed a very low Po under normal recording conditions in the absence of CaMKII, thus mimicking a channel that has been phosphorylated by CaMKII. The results of this study show for the first time that Serine-150 of the …
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Functional significance of Hsp70 post-translational modification in prion propagation and cellular function
… assessed a variety of non-phosphorylateable and phosphomimetic Hsp70 mutants for phenotypic alterations in Hsp70 functions. It was found that alteration of Hsp70 phosphorylation status in vivo can impair prion propagation, alter both basal and acquired thermotolerance and in some cases render …
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Dissecting the Physiological Roles of ULK1/2 in the Mouse Brain
… were reversed upon expression of SEC16A with a phosphomimetic substitution. Thus, the regulation of ER-to-Golgi trafficking by ULK1/2 is essential for cellular homeostasis. Moreover, the defect in SERT trafficking may also contribute to the disrupted formation of the barrel cortex in the Ulk1/2 …
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