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Cited by 308Cytokine Signaling Pathways and Interactionsinterferon and immune responsesGenomics and Chromatin Dynamics

DNA binding of in vitro activated Stat1 alpha, Stat1 beta and truncated Stat1: interaction between NH2‐terminal domains stabilizes binding of two dimers to tandem DNA sites.

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Vinkemeier, U., Cohen, S. L., Moarefi, I., Chait, B. T., Kuriyan, J., Darnell, J. E.·1996·The EMBO Journal
DOI 10.1002/j.1460-2075.1996.tb00946.xPMC PMC452306
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Abstract

Stat1 alpha, Stat1 beta and a proteolytically defined truncated Stat1 (132-713, Stat1tc) have been prepared from recombinant sources. All three proteins were specifically phosphorylated on Tyr701 in vitro and the phosphoprotein purified to homogeneity. This was achieved by employing a new isolation scheme that does not include DNA affinity steps and readily allows for the isolation of tens of milligrams of activated Stat protein. The purified phosphoprotein was free of traces of unphosphorylated polypeptide as detected by mass spectrometry. The phosphorylated Stat1 preparations bound to various DNA recognition sites with the same Keq of approximately 1 x 10(-9) M; distinction between 'weak' and 'strong' binding sites is determined by the very rapid dissociation (< 30 s, t1/2) from 'weak' sites compared with 'strong' sites (approximately 3 min, t1/2). Reports of 'weak' tandem binding sites in a natural gene caused us to examine binding to tandem sites leading to the finding that the Stat1 alpha or beta (38 amino acids shorter on the C terminus) bound to two tandem sites (but not two head-to-head sites) with a higher stability than to a single recognition site. The N-terminally truncated protein Stat1tc did not show this cooperative binding, thus implicating the N-terminal domain in promoting Stat1-Stat1 dimer interaction.

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Crossref · 10.1002/j.1460-2075.1996.tb00946.x

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  • Despite having similar apparent Kd values, the binding with DNA may differ significantly in rates of association with and dissociation from the Stat1 protein.

    “Quantitative analysis of the Stat1-DNA exchange reaction had revealed identical affinities for various sites, but despite having similar apparent K d values, the binding with DNA may differ significantly in rates of association with and dissociation from the Stat1 protein (Vinkemeier et al. 1996).”

    From DNA binding controls inactivation and nuclear accumulation of the transcription factor Stat1 (2003) · cites this paper as Vinkemeier et al. 1996

  • Quantitative analysis of the Stat1-DNA exchange reaction had revealed identical affinities for various sites.

    “Quantitative analysis of the Stat1-DNA exchange reaction had revealed identical affinities for various sites, but despite having similar apparent K d values, the binding with DNA may differ significantly in rates of association with and dissociation from the Stat1 protein (Vinkemeier et al. 1996).”

    From DNA binding controls inactivation and nuclear accumulation of the transcription factor Stat1 (2003) · cites this paper as Vinkemeier et al. 1996

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References 32 · Cited by 308

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Authors
Vinkemeier, U., Cohen, S. L., Moarefi, I., Chait, B. T., Kuriyan, J., & Darnell, J. E.
Affiliations
Rockefeller University (US)
ORCID
Venue
The EMBO Journal
Publisher
Springer Science and Business Media LLC
Published
1996-10-01
Volume
15
Issue
20
Pages
5616-5626
ISSN
  • 0261-4189 (ISSN-L)
  • 1460-2075
DOI
10.1002/j.1460-2075.1996.tb00946.x
PMC
PMC452306
OpenAlex
W103687069
Cited by
308
References
32
Citations by year
2026: 5 · 2025: 5 · 2024: 3 · 2023: 8 · 2022: 7 · 2021: 5 · 2020: 5 · 2019: 5 · 2018: 5 · 2017: 4 · 2016: 5 · 2015: 6 · 2014: 10 · 2013: 9 · 2012: 15
FWCI
8.69
Type
journal article
Language
en
Open access
Licence
Locations
  • The EMBO Journal (publishedVersion)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institute of Allergy and Infectious Diseases (AI 34420, R01AI032489, AI32489)
  • National Center for Research Resources (RR00862, P41RR000862)
Topics
  • Health Sciences › Medicine › Oncology › Cytokine Signaling Pathways and Interactions
  • Life Sciences › Immunology and Microbiology › Immunology › interferon and immune responses
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Genomics and Chromatin Dynamics
Keywords
STAT1, DNA binding, cooperative DNA binding, N-terminal domain, protein-DNA interactions, interferon-stimulated genes
MeSH terms
Binding Sites, DNA, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Humans, Kinetics, Protein Conformation, Recombinant Proteins, Transcription Factors, Transfection, Signal Transduction, Trans-Activators, DNA, Complementary, STAT1 Transcription Factor, Interferon-Stimulated Gene Factor 3, DNA Fragmentation, ErbB Receptors

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