Corrigendum: Notch Intracellular Domain Plasmid Delivery via Poly(Lactic-Co-Glycolic Acid) Nanoparticles to Upregulate Notch Pathway Molecules

Notch signaling is a highly conserved signaling system that is required for embryonic development and regeneration of organs. When the signal is lost, maldevelopment occurs and leads to a lethal state. Delivering exogenous genetic materials encoding Notch into cells can reestablish downstream signaling and rescue cellular functions. In this study, we utilized the negatively charged and FDA approved polymer poly(lactic-co-glycolic acid) to encapsulate Notch Intracellular Domain-containing plasmid in nanoparticles. We show that primary human umbilical vein endothelial cells (HUVECs) readily uptake the nanoparticles with and without specific antibody targets. We demonstrated that our nanoparticles are non-toxic, stable over time, and compatible with blood. We further demonstrated that HUVECs could be successfully transfected with these nanoparticles in static and dynamic environments. Lastly, we elucidated that these nanoparticles could upregulate the downstream genes of Notch signaling, indicating that the payload was viable and successfully altered the genetic downstream effects.

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The Notch Intracellular Domain Can Function as a Coactivator for LEF-1

Molecular and Cellular Biology ◽

10.1128/mcb.21.22.7537-7544.2001 ◽

2001 ◽

Vol 21 (22) ◽

pp. 7537-7544 ◽

Cited By ~ 75

Author(s):

David A. Ross ◽

Tom Kadesch

Keyword(s):

Notch Signaling ◽

Wnt Signaling Pathway ◽

Notch Pathway ◽

High Mobility ◽

In Vitro Assays ◽

Transactivation Domain ◽

Intracellular Domain ◽

Notch Intracellular Domain

ABSTRACT Notch signaling commences with two ligand-mediated proteolysis events that release the Notch intracellular domain, NICD, from the plasma membrane. NICD then translocates into the nucleus and interacts with the DNA binding protein CSL to activate transcription. We found that NICD expression also potentiates activity of the transcription factor LEF-1. NICD stimulation of LEF-1 activity was context dependent and occurred on a subset of promoters distinct from those activated by β-catenin. Importantly, the effect of NICD does not appear to be mediated through canonical components of the Wnt signaling pathway or downstream components of the Notch pathway. In vitro assays show a weak association between the C-terminal transactivation domain of NICD and the high-mobility group domain of LEF-1, suggesting that the two proteins interact in vivo. Our data therefore describe a new nuclear target of Notch signaling and a new coactivator for LEF-1.

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Notch Intracellular Domain Plasmid Delivery via Poly(lactic-co-glycolic acid) Nanoparticles to Upregulate Notch Signaling

10.1101/2021.04.16.440241 ◽

2021 ◽

Author(s):

Victoria Messerschmidt ◽

Aneetta Kuriakose ◽

Uday Chintapula ◽

Samantha Laboy-Segarra ◽

Thuy Truong ◽

...

Keyword(s):

Endothelial Cells ◽

Notch Signaling ◽

Glycolic Acid ◽

Umbilical Vein ◽

Genetic Material ◽

Human Umbilical Vein ◽

Notch Intracellular Domain ◽

Downstream Effects ◽

Umbilical Vein Endothelial Cells ◽

Plasmid Delivery

Notch signaling is a highly conserved signaling system that is required for embryonic development and regeneration of organs. When the signal is lost, maldevelopment occurs and leads to a lethal state. Liposomes and retroviruses are most commonly used to deliver genetic material to cells. However, there are many drawbacks to these systems such as increased toxicity, nonspecific delivery, short half-life, and stability after formulation. We utilized the negatively charged and FDA approved polymer poly(lactic-co-glycolic acid) to encapsulate Notch Intracellular Domaincontaining plasmid in nanoparticles. In this study, we show that primary human umbilical vein endothelial cells readily uptake the nanoparticles with and without specific antibody targets. We demonstrated that our nanoparticles also are nontoxic, stable over time, and compatible with blood. We also determined that we can successfully transfect primary human umbilical vein endothelial cells (HUVECs) with our nanoparticles in static and dynamic environments. Lastly, we elucidated that our transfection upregulates the downstream genes of Notch signaling, indicating that the payload was viable and successfully altered the genetic downstream effects.

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Notch family members follow stringent requirements for intracellular domain dimerization at sequence-paired sites

10.1101/2020.05.20.106054 ◽

2020 ◽

Author(s):

Jacob J. Crow ◽

Allan R. Albig

Keyword(s):

Transcriptional Activation ◽

Target Genes ◽

Regulatory Mechanism ◽

Family Members ◽

Regulatory Function ◽

Specific Nature ◽

Intracellular Domain ◽

Notch Intracellular Domain ◽

Highly Sensitive ◽

Cellular Identity

ABSTRACTNotch signaling is essential for multicellular life, regulating core functions such as cellular identity, differentiation, and fate. These processes require highly sensitive systems to avoid going awry, and one such regulatory mechanism is through Notch intracellular domain dimerization. Select Notch target genes contain sequence-paired sites (SPS); motifs in which two Notch transcriptional activation complexes can bind and interact through Notch’s ankyrin domain, resulting in enhanced transcriptional activation. This mechanism has been mostly studied through Notch1, and to date, the abilities of the other Notch family members have been left unexplored. Through the utilization of minimalized, SPS-driven luciferase assays, we were able to test the functional capacity of Notch dimers. Here we show that each family member is capable of dimerization-induced signaling, following the same stringent requirements as seen with Notch1. Interestingly, we identified a mechanical difference between canonical and cryptic SPSs, leading to differences in their dimerization-induced regulation. Finally, we profiled the Notch family members’ SPS gap distance preferences and found that they all prefer a 16-nucleotide gap, with little room for variation. In summary, this work highlights the potent and highly specific nature of Notch dimerization and refines the scope of this regulatory function.

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Degradation of the NOTCH intracellular domain by elevated autophagy in osteoblasts promotes osteoblast differentiation and alleviates osteoporosis

Autophagy ◽

10.1080/15548627.2021.2017587 ◽

2022 ◽

pp. 1-10

Author(s):

Gota Yoshida ◽

Tsuyoshi Kawabata ◽

Hyota Takamatsu ◽

Shotaro Saita ◽

Shuhei Nakamura ◽

...

Keyword(s):

Osteoblast Differentiation ◽

Intracellular Domain ◽

Notch Intracellular Domain

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RHOQ is induced by DLL4 and regulates angiogenesis by determining the intracellular route of the Notch intracellular domain

Angiogenesis ◽

10.1007/s10456-020-09726-w ◽

2020 ◽

Vol 23 (3) ◽

pp. 493-513

Author(s):

Esther Bridges ◽

Helen Sheldon ◽

Esther Kleibeuker ◽

Evelyn Ramberger ◽

Christos Zois ◽

...

Keyword(s):

Intracellular Domain ◽

Notch Intracellular Domain

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The Notch signalling pathway and breast cancer: The importance of balance and cellular self-control.

Current Signal Transduction Therapy ◽

10.2174/1574362414666190916120659 ◽

2019 ◽

Vol 14 ◽

Author(s):

Germán Saucedo-Correa ◽

Alejandro Bravo-Patiño ◽

Rosa Elvira Núñez-Anita ◽

Javier Oviedo-Boyso ◽

Juan José Valdez-Alarcón ◽

...

Keyword(s):

Notch Signaling ◽

Signaling Pathway ◽

Cross Talk ◽

Transcriptional Activation ◽

Notch Pathway ◽

Notch Signaling Pathway ◽

Self Control ◽

Design Strategies ◽

Notch Intracellular Domain ◽

The Cross

Notch is a cell-signaling pathway that is highly conserved in all metazoans and is responsible for cell differentiation and cross-talk communication with other signaling pathways such as WNT and Hh. In most cancers, the Notch signaling pathway is altered, causing atypical activity of vital processes such as cell cycle, differentiation and apoptosis, leading the cell to a carcinogenic state. Currently, the Notch signaling pathway has taken a special interest to design strategies in order to regulate the activity of this pathway since it is known that in the cancer molecular micro-environment the Notch pathway is over-expressed or presents an aberrant function, which, in consequence, corrupts the cross-talk communication with WNT and Hh pathways. Most of the existing strategies are focused on the systematic and whole inhibition of Notch pathway at the membrane level by the use of γ-secretases inhibitors. There are few strategies that act at the nuclear level inhibiting the activity of the transcriptional activation complex composed by the Notch intracellular domain, the transcriptional factor CSL and the Mastermind co-activator. In this review, by the fact that there are not any strategy focused to revert the over expression effect caused by the Notch pathway constitutive activity, we propose that the efforts to develop new strategies against cancer should be focused to understand the complexity of the cross-talk communication between Notch, WNT and Hh pathways to neutralize the gene aberrant activity characteristic of cancer cells which are responsible for those corrupted cross-talk communication.

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Extension of the Notch intracellular domain ankyrin repeat stack by NRARP promotes feedback inhibition of Notch signaling

Science Signaling ◽

10.1126/scisignal.aay2369 ◽

2019 ◽

Vol 12 (606) ◽

pp. eaay2369 ◽

Cited By ~ 3

Author(s):

Sanchez M. Jarrett ◽

Tom C. M. Seegar ◽

Mark Andrews ◽

Guillaume Adelmant ◽

Jarrod A. Marto ◽

...

Keyword(s):

Notch Signaling ◽

Negative Feedback ◽

Transcriptional Activation ◽

Cultured Cells ◽

Feedback Inhibition ◽

Lymphoblastic Leukemia ◽

Ankyrin Repeat ◽

Structural Basis ◽

Intracellular Domain ◽

Notch Intracellular Domain

Canonical Notch signaling relies on regulated proteolysis of the receptor Notch to generate a nuclear effector that induces the transcription of Notch-responsive genes. In higher organisms, one Notch-responsive gene that is activated in many different cell types encodes the Notch-regulated ankyrin repeat protein (NRARP), which acts as a negative feedback regulator of Notch responses. Here, we showed that NRARP inhibited the growth of Notch-dependent T cell acute lymphoblastic leukemia (T-ALL) cell lines and bound directly to the core Notch transcriptional activation complex (NTC), requiring both the transcription factor RBPJ and the Notch intracellular domain (NICD), but not Mastermind-like proteins or DNA. The crystal structure of an NRARP-NICD1-RBPJ-DNA complex, determined to 3.75 Å resolution, revealed that the assembly of NRARP-NICD1-RBPJ complexes relied on simultaneous engagement of RBPJ and NICD1, with the three ankyrin repeats of NRARP extending the Notch1 ankyrin repeat stack. Mutations at the NRARP-NICD1 interface disrupted entry of the proteins into NTCs and abrogated feedback inhibition in Notch signaling assays in cultured cells. Forced expression of NRARP reduced the abundance of NICD in cells, suggesting that NRARP may promote the degradation of NICD. These studies establish the structural basis for NTC engagement by NRARP and provide insights into a critical negative feedback mechanism that regulates Notch signaling.

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