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Related Concept Videos

Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Phosphorylation01:02

Phosphorylation

The addition or removal of phosphate groups from proteins is the most common chemical modification that regulates cellular processes. These modifications can affect the structure, activity, stability, and localization of proteins within cells as well as their interactions with other proteins.
During phosphorylation, protein kinases transfer the terminal phosphate group of ATP to specific amino acid side chains of substrate proteins. Serine, threonine, and tyrosine are the most commonly...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Protein Kinases and Phosphatases02:54

Protein Kinases and Phosphatases

Proteins undergo chemical modifications that trigger changes in the charge, structure, and conformation of the proteins. Phosphorylation, acetylation, glycosylation, nitrosylation, ubiquitination, lipidation, methylation, and proteolysis are various protein modifications that regulate protein activity. Such modifications are usually enzyme-driven.
Protein kinases
Many proteins in the cell are regulated by phosphorylation, the addition of a phosphate group. A family of enzymes called kinases...
Somatic to iPS Cell Reprogramming01:29

Somatic to iPS Cell Reprogramming

Reprogramming alters the gene expression in somatic cells, transforming them into induced pluripotent stem (iPS) cells over several generations. Scientists can reprogram cells by introducing genes for four transcription factors—Oct4, Sox2, Klf4, and c-Myc (OSKM) by viral or non-viral methods. These factors are also known as Yamanaka factors after Shinya Yamanaka, who first generated iPS cells using mouse skin cells. Yamanaka was awarded the Nobel Prize in Physiology or Medicine in 2012 for this...
Methods of Nuclear Reprogramming01:24

Methods of Nuclear Reprogramming

Nuclear reprogramming is a process of transforming one cell type into an unrelated cell type by epigenetic changes that alter the cell’s original gene expression pattern. Such epigenetic changes force cells to express a different set of genes, which play a significant role in inducing transformation into other cell types. Nuclear reprogramming offers applications in reproductive cloning for livestock propagation and regenerative medicine — developing patient-specific cells for injury repair.

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Related Experiment Video

Updated: May 23, 2026

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
08:01

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal

Published on: May 30, 2012

Phosphorylation regulates human OCT4.

Justin Brumbaugh1, Zhonggang Hou, Jason D Russell

  • 1Department of Biomolecular Chemistry, University of Wisconsin, Madison, WI 53706-1532, USA.

Proceedings of the National Academy of Sciences of the United States of America
|April 5, 2012
PubMed
Summary
This summary is machine-generated.

Researchers identified new phosphorylation sites on OCT4 (octamer-binding transcription factor 4), a key protein for pluripotency. Phosphorylation at T234/S235 negatively impacts OCT4 DNA binding and function, linking ERK signaling to pluripotency regulation.

More Related Videos

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Related Experiment Videos

Last Updated: May 23, 2026

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal
08:01

Oct4GiP Reporter Assay to Study Genes that Regulate Mouse Embryonic Stem Cell Maintenance and Self-renewal

Published on: May 30, 2012

Oligopeptide Competition Assay for Phosphorylation Site Determination
09:16

Oligopeptide Competition Assay for Phosphorylation Site Determination

Published on: May 18, 2017

Area of Science:

  • Stem cell biology
  • Molecular and cellular biology
  • Epigenetics

Background:

  • Octamer-binding transcription factor 4 (OCT4) is crucial for maintaining embryonic stem cell pluripotency and self-renewal.
  • Understanding post-translational modifications, such as phosphorylation, is key to elucidating OCT4's regulatory mechanisms.

Purpose of the Study:

  • To identify and characterize novel phosphorylation sites on OCT4.
  • To investigate the functional impact of OCT4 phosphorylation on its DNA-binding activity and role in cellular reprogramming.
  • To explore the relationship between OCT4 phosphorylation and signaling pathways, particularly ERK signaling.

Main Methods:

  • Liquid chromatography-mass spectrometry (LC-MS) was employed to identify phosphosites on OCT4 and its interacting partners.
  • Functional assays, including reporter gene assays and cellular reprogramming experiments, were used to assess the impact of specific phosphorylation sites.
  • Kinase assays were performed to confirm direct phosphorylation by ERK2.

Main Results:

  • 14 localized phosphorylation sites on OCT4 were identified, 11 of which are novel.
  • Phosphorylation at T234 and S235 within the homeobox region was found to negatively regulate OCT4's DNA-binding activity and transcriptional function.
  • Mutating T234/S235 to mimic constitutive phosphorylation decreased reporter gene activation and reprogramming efficiency.
  • 144 unique phosphopeptides were identified on OCT4-interacting proteins (e.g., SOX2, SALL4), enriched for ERK signaling motifs.
  • ERK2 was confirmed to directly phosphorylate OCT4 at specific sites in vitro.

Conclusions:

  • Phosphorylation of OCT4, particularly at T234/S235, acts as a negative regulator of its function in maintaining pluripotency.
  • The study establishes a direct mechanistic link between ERK signaling and the regulation of pluripotency by OCT4 and its associated factors.