The intracellular domain of mouse Notch: a constitutively activated repressor of myogenesis directed at the basic helix-loop-helix region of MyoD

Development ◽  
1994 ◽  
Vol 120 (9) ◽  
pp. 2385-2396 ◽  
Author(s):  
R. Kopan ◽  
J.S. Nye ◽  
H. Weintraub
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Cultured Cells ◽  
Ectopic Expression ◽  
Presomitic Mesoderm ◽  
Intracellular Domain ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Localization Signal ◽  
Cell Commitment

We show that Myf-5 and mNotch mRNA are both present in the presomitic mesoderm before muscle cell commitment and before muscle structural gene activation. The failure of presomitic mesoderm to respond to Myf-5 and express myogenic properties implies that there may be a mechanism in presomitic mesoderm to suppress muscle differentiation. Here we show that ectopic expression of the intracellular domain of mNotch (mNotchIC) functions as a constitutively activated repressor of myogenesis both in cultured cells and in frog embryos. Mutagenesis experiments indicate that the target for inactivation by mNotch is the MyoD basic helix-loop-helix domain. mNotchIC contains a nuclear localization signal and localizes to the nucleus. Removal of the nuclear localization signal (NLS) reduces nuclear localization and diminishes the inhibition of myogenesis caused by Myf-5 or MyoD. Additional experiments show that the CDC10/SWI6/ankyrin repeats are also necessary for myogenic inhibition.

scholarly journals ADAM10- and presenilin 1/g-secretase-dependent cleavage of PTPRT mitigates Neurodegeneration of Alzheimer’s disease

2021 ◽  
Author(s):  
Siling Liu ◽  
Zhongyu Zhang ◽  
Lianwei Li ◽  
Li Yao ◽  
Zhanshan Ma ◽  
...  
Keyword(s):  
Alzheimer’S Disease ◽  
Alzheimer's Disease ◽  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Transmembrane Protein ◽  
Neuronal Cell ◽  
Synaptic Function ◽  
Presenilin 1 ◽  
Intracellular Domain ◽  
Localization Signal

Abstract BackgroundPTPRT (receptor-type tyrosine-protein phosphatase T), as a brain-specific type 1 transmembrane protein, plays an important function in neurodevelopment and synapse formation. However, whether PTPRT-dependent signaling is involved in Alzheimer’s disease (AD) remains elusive. MethodsWe have begun to address these questions by investigation of the relationship between Ab and PTPRT-dependent signaling in both postmortem human AD brain tissues, APP/PS1 mice, and different cell lines using biochemical and histological methods. Using RNAseq we analyzed gene expression regulated by PTPRT intracellular domain (PICD). We further assessed the function of PICD in AD brain using neuropathology, electrophysiology and animal behavior testing after conduction on APP/PS1 mice with overexpressing PICD in the hippocampus. ResultsA significant downregulation in the levels of Ptprt mRNA and protein was found in both human AD and disease model mouse brains. PTPRT intracellular domain (PICD), which was released from ADAM10- and presenilin 1-/g-secretase-dependent cleavage of PTPRT, efficiently translocated to the nucleus via a conserved nuclear localization signal. Inhibition of nuclear-localization of PICD via the mutation of its nuclear localization signal (NLS) leads to accumulation of phosphorylated signal transducer and activator of transcription 3 (pSTAT3), which is a substrate of PTPRT and eventually resulted in neuronal cell death. RNA sequencing reveals that expression of the PICD alone can profoundly alter the expression of genes associated with synapse function and dephosphorylation et al. Overexpression of PICD alone not only significantly decreases the level of phosph-STAT3Y705 and Ab deposition in the hippocampus of APP/PS1 mice, but also improves synaptic function and behavioral deficits in APP/PS1 mice. ConclusionsAltogether, these findings suggest that a novel role of the ADAM 10- and presenilin 1-/g-secretase-dependent cleavage of PTPRT in the events can mitigate neurodegeneration of AD and moderate Alzheimer’s pathogenesis.

scholarly journals Acetylation of nuclear localization signal controls importin-mediated nuclear transport of Ku70

2018 ◽  
Author(s):  
H Fujimoto ◽  
T Ikuta ◽  
A Koike ◽  
M Koike
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Cultured Cells ◽  
Nuclear Transport ◽  
Intracellular Localization ◽  
Lysine Residues ◽  
Importin Α ◽  
Localization Signal ◽  
Transport Factors

AbstractKu70 participates in various intra-and extra-nucleic processes. For multifunctional control, machinery that precisely regulates the intracellular localization of Ku70 is essential. Recently, it was reported that acetylation of Ku70 regulates its function. Here, we demonstrate that specific lysine residues in Ku70 that are targets of acetylation are critical for regulating nuclear transport in vivo. Ku70-GFP fusion proteins transiently expressed in cultured cells localized in the nucleus, whereas mimicking acetylation of K553 or K556 in the Ku70 nuclear localization signal (NLS) by substituting these lysine residues with glutamine markedly decreased the nuclear localization of Ku70. Moreover, the Ku70-importin interaction was suppressed in the K553Q and K556Q mutants. Theoretical estimations indicated that the binding energy between the Ku70 NLS and importin-α decreases with acetylation of lysine residues in the Ku70 NLS, similar to the case when these lysine residues are substituted with glutamine. These results suggest that acetylation of specific lysine residues in the Ku70 NLS is a key switch that controls the localization of Ku70 by modulating interactions between Ku70 and nuclear transport factors.

scholarly journals Regulation of bHLH-PAS protein subcellular localization during Drosophila embryogenesis

Development ◽  
1998 ◽  
Vol 125 (9) ◽  
pp. 1599-1608 ◽  
Author(s):  
M.P. Ward ◽  
J.T. Mosher ◽  
S.T. Crews
Keyword(s):  
Subcellular Localization ◽  
Nuclear Localization ◽  
Ectopic Expression ◽  
Nuclear Accumulation ◽  
Salivary Duct ◽  
Cell Lineages ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix ◽  
Cns Midline ◽  
Midline Cells

The Drosophila Single-minded and Tango basic-helix-loop-helix-PAS protein heterodimer controls transcription and embryonic development of the CNS midline cells, while the Trachealess and Tango heterodimer controls tracheal cell and salivary duct transcription and development. Expression of both single-minded and trachealess is highly restricted to their respective cell lineages, however tango is broadly expressed. The developmental control of subcellular localization of these proteins is investigated because of their similarity to the mammalian basic-helix-loop-helix-PAS Aromatic hydrocarbon receptor whose nuclear localization is dependent on ligand binding. Confocal imaging of Single-minded and Trachealess protein localization indicate that they accumulate in cell nuclei when initially synthesized in their respective cell lineages and remain nuclear throughout embryogenesis. Ectopic expression experiments show that Single-minded and Trachealess are localized to nuclei in cells throughout the ectoderm and mesoderm, indicating that nuclear accumulation is not regulated in a cell-specific fashion and unlikely to be ligand dependent. In contrast, nuclear localization of Tango is developmentally regulated; it is localized to the cytoplasm in most cells except the CNS midline, salivary duct, and tracheal cells where it accumulates in nuclei. Genetic and ectopic expression experiments indicate that Tango nuclear localization is dependent on the presence of a basic-helix-loop-helix-PAS protein such as Single-minded or Trachealess. Conversely, Drosophila cell culture experiments show that Single-minded and Trachealess nuclear localization is dependent on Tango since they are cytoplasmic in the absence of Tango. These results suggest a model in which Single-minded and Trachealess dimerize with Tango in the cytoplasm of the CNS midline cells and trachea, respectively, and the dimeric complex accumulates in nuclei in a ligand-independent mode and regulates lineage-specific transcription. The lineage-specific action of Single-minded and Trachealess derives from transcriptional activation of their genes in their respective lineages, not from extracellular signaling.

scholarly journals Reactivation of Epstein–Barr virus can be triggered by an Rta protein mutated at the nuclear localization signal

2005 ◽  
Vol 86 (2) ◽  
pp. 317-322 ◽  
Author(s):  
Tsuey-Ying Hsu ◽  
Yao Chang ◽  
Pei-Wen Wang ◽  
Mei-Ying Liu ◽  
Mei-Ru Chen ◽  
...  
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Epstein Barr Virus ◽  
Target Genes ◽  
Ectopic Expression ◽  
Epithelial Cell Line ◽  
Barr Virus ◽  
Early Protein ◽  
Localization Signal ◽  
Epstein Barr

Rta, an immediate-early protein of Epstein–Barr virus (EBV), is a transcriptional activator that induces lytic gene expression and triggers virus reactivation. Being located predominantly in the nucleus, Rta can exert its transactivation function through either direct DNA binding or certain indirect mechanisms mediated by cellular signalling and other transcriptional factors. This study examined whether the subcellular localization of Rta was critical for the induction of target genes. First, 410KRKK413 was identified as a nuclear localization signal (NLS) of Rta. An Rta mutant with the NLS converted to 410AAAA413 showed cytoplasmic localization and failed to activate the promoter of BGLF5. Interestingly, ectopic expression of the Rta mutant still disrupted EBV latency in an epithelial cell line. Reporter gene assays revealed that the NLS-mutated Rta retained the ability to activate two lytic promoters, Zp and Rp, at a considerable level. Thus, the cytoplasmic Rta mutant could induce expression of endogenous Zta and Rta, triggering reactivation of EBV.

scholarly journals Coordinate Nuclear Targeting of the FANCD2 and FANCI Proteins via a FANCD2 Nuclear Localization Signal

PLoS ONE ◽  
2013 ◽  
Vol 8 (11) ◽  
pp. e81387 ◽  
Author(s):  
Rebecca A. Boisvert ◽  
Meghan A. Rego ◽  
Paul A. Azzinaro ◽  
Maurizio Mauro ◽  
Niall G. Howlett
Keyword(s):  
Nuclear Localization ◽  
Nuclear Localization Signal ◽  
Nuclear Targeting ◽  
Localization Signal
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