Structural and mechanistic insights into nuclear transport and delivery of the critical pluripotency factor Oct4 to DNA

Okuyama, Takahide; Yamagishi, Ryosuke; Shimada, Jiro; Ikeda, Masa‐Aki; Maruoka, Yayoi; Kaneko, Hiroki

doi:10.1080/07391102.2017.1289124

Cited by 3 publications

(3 citation statements)

References 46 publications

(69 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…OCT4 is transported through nuclear pores by the nuclear export protein CRM1 (also known as XPO1) [20,24] and the nuclear import protein, importin α [25][26][27]. Therefore, we investigated the involvement of CRM1 in the translocation of pluripotency-related transcription factors in human ESCs during uniaxial forces.…”

Section: Resultsmentioning

confidence: 99%

Rapid translocation of pluripotency-related transcription factors by external uniaxial forces

Topal

Kim

Villa‐Diaz

et al. 2019

Integrative Biology

View full text Add to dashboard Cite

Human embryonic stem cells subjected to a one-time uniaxial stretch for as short as 30-min on a flexible substrate coated with Matrigel experienced rapid and irreversible nuclear-to-cytoplasmic translocation of NANOG and OCT4, but not Sox2. Translocations were directed by intracellular transmission of biophysical signals from cell surface integrins to nuclear CRM1 and were independent of exogenous soluble factors. On E-CADHERIN-coated substrates, presumably with minimal integrin engagement, mechanical strain-induced rapid nuclear-to-cytoplasmic translocation of the three transcription factors. These findings might provide fundamental insights into early developmental processes and may facilitate mechanotransductionmediated bioengineering approaches to influencing stem cell fate determination. Insight, innovation, integration How do mechanical stimuli, sensed through different types of cell adhesions, alter embryonic stem cell fate? This paper employs a stretchable device coated with different cell adhesion molecules to apply stretch to human embryonic stem cells in a manner that might mimic differentiation of cell lineages present at the blastocyst stage of development. The work reveals that stretch induces an unexpectedly rapid export of a subset of pluripotent transcription factors from the nucleus to the cytoplasm before significant decreases in overall levels. Interestingly, the subset of transcription factors exported differs between cells stretched on Matrigel-versus E-CADHERIN-coated devices. These results have physiological relevance to embryonic development where some cells experience mechanical forces predominantly through cell-cell contact while other cells also experience cell-ECM interactions leading to different cell-fate specification. These insights might enhance our understanding of early development and may also guide bioengineering approaches where stretching is used to accelerate or regulate the direction of differentiation.

show abstract

Section: Resultsmentioning

confidence: 99%

Rapid translocation of pluripotency-related transcription factors by external uniaxial forces

Topal

Kim

Villa‐Diaz

et al. 2019

Integrative Biology

View full text Add to dashboard Cite

show abstract

“…Furthermore, the NLS fragment from the critical pluripotency factor Oct4 adopts a bound pose in the impα major site, which is inconsistent with that of VEEV NLS. For example, P5 Arg escapes the cage formed by the tryptophan side chains . In contrast to these NLS fragments, the VEEV coreNLS peptide retains native-like binding pose in the major NLS binding site .…”

Section: Resultsmentioning

confidence: 99%

“…For example, P5 Arg escapes the cage formed by the tryptophan side chains. 36 In contrast to these NLS fragments, the VEEV coreNLS peptide retains native-like binding pose in the major NLS binding site. 15 It is notable that none of these other NLS sequences harbor the VEEV minNLS sequence KKPK.…”

Section: Resultsmentioning

confidence: 99%

Competitive Binding of Viral Nuclear Localization Signal Peptide and Inhibitor Ligands to Importin-α Nuclear Transport Protein

Delfing,

Laracuente,

Jeffries

et al. 2024

J. Chem. Inf. Model.

View full text Add to dashboard Cite

Venezuelan equine encephalitis virus (VEEV) is a highly virulent pathogen whose nuclear localization signal (NLS) sequence from capsid protein binds to the host importin-α transport protein and blocks nuclear import. We studied the molecular mechanisms by which two small ligands, termed I1 and I2, interfere with the binding of VEEV's NLS peptide to importin-α protein. To this end, we performed all-atom replica exchange molecular dynamics simulations probing the competitive binding of the VEEV coreNLS peptide and I1 or I2 ligand to the importin-α major NLS binding site. As a reference, we used our previous simulations, which examined noncompetitive binding of the coreNLS peptide or the inhibitors to importin-α. We found that both inhibitors completely abrogate the native binding of the coreNLS peptide, forcing it to adopt a manifold of nonnative loosely bound poses within the importin-α major NLS binding site. Both inhibitors primarily destabilize the native coreNLS binding by masking its amino acids rather than competing with it for binding to importin-α. Because I2, in contrast to I1, binds off-site localizing on the edge of the major NLS binding site, it inhibits fewer coreNLS native binding interactions than I1. Structural analysis is supported by computations of the free energies of the coreNLS peptide binding to importin-α with or without competition from the inhibitors. Specifically, both inhibitors reduce the free energy gain from coreNLS binding, with I1 causing significantly larger loss than I2. To test our simulations, we performed AlphaScreen experiments measuring IC50 values for both inhibitors. Consistent with in silico results, the IC50 value for I1 was found to be lower than that for I2. We hypothesize that the inhibitory action of I1 and I2 ligands might be specific to the NLS from VEEV's capsid protein.

show abstract

Insights into the Giardia intestinalis enolase and human plasminogen interaction

et al. 2017

View full text Add to dashboard Cite

Giardia intestinalis is an intestinal parasite that causes diarrhea in humans and animals worldwide. The enolase of G. intestinalis (GiENO) participates in its glycolysis pathway and is abundantly expressed in the parasite cytosol; however, its localization on the surface of trophozoites and cysts has been demonstrated. Enolases from bacteria and parasites can have different functions and are considered moonlighting proteins, for example, as a cell surface plasminogen receptor. In relation to GiENO, no studies have been performed about its possible participation as a plasminogen receptor. In this work, we employed molecular docking and multiscale molecular dynamics (MD) simulations to explore the possible interactions of human plasminogen (HsPLG) with the open and closed GiENO conformations. Our proposed GiENO plasminogen binding site (PLGBs) was identified at Lys266 based on the sequence comparison with bacterial enolase known to act as a plasminogen receptor. Our docking results performed with multiple MD snapshots of the closed GiENO conformation showed that Lys266 preferentially binds to the K5 domain of HsPLG. On the other hand, open GiENO conformations from all-atom and coarse-grained simulations indicated a high preference of the HsPLG K4 domain for lysine residues 186 and 188. Furthermore, we identified a potential N-glycosylation site of GiENO which suggests a possible explanation for the parasite cell surface localization or host mucin oligosaccharide adhesion mechanism. Our study constitutes the first multiscale computational study to explore the plasminogen receptor function of GiENO for its further consideration as a potential therapeutic target for giardiasis treatment.

show abstract

Structural and mechanistic insights into nuclear transport and delivery of the critical pluripotency factor Oct4 to DNA

Cited by 3 publications

References 46 publications

Rapid translocation of pluripotency-related transcription factors by external uniaxial forces

Rapid translocation of pluripotency-related transcription factors by external uniaxial forces

Competitive Binding of Viral Nuclear Localization Signal Peptide and Inhibitor Ligands to Importin-α Nuclear Transport Protein

Insights into the Giardia intestinalis enolase and human plasminogen interaction

Contact Info

Product

Resources

About