Real time analysis of STAT3 nucleocytoplasmic shuttling
Open accessAbstract
The transcription factor STAT3 is most important for the signal transduction of interleukin-6 and related cytokines. Upon stimulation cytoplasmic STAT3 is phosphorylated at tyrosine 705, translocates into the nucleus, and induces target genes. Notably, STAT proteins are also detectable in the nuclei of unstimulated cells. In this report we introduce a new method for the real time analysis of STAT3 nucleocytoplasmic shuttling in living cells which is based on the recently established fluorescence localization after photobleaching (FLAP) approach. STAT3 was C-terminally fused with the cyan (CFP) and yellow (YFP) variants of the green fluorescent protein. In the resulting STAT3-CFP-YFP (STAT3-CY) fusion protein the YFP can be selectively bleached using the 514-nm laser of a confocal microscope. This setting allows studies on the dynamics of STAT3 nucleocytoplasmic transport by monitoring the subcellular distribution of fluorescently labeled and selectively bleached STAT3-CY. By this means we demonstrate that STAT3-CY shuttles continuously between the cytosol and the nucleus in unstimulated cells. This constitutive shuttling does not depend on the phosphorylation of tyrosine 705 because a STAT3(Y705F)-CY mutant shuttles to the same extent as STAT3-CY. Experiments with deletion mutants reveal that the N-terminal moiety of STAT3 is essential for shuttling. Further studies suggest that a decrease in STAT3 nuclear export contributes to the nuclear accumulation of STAT3 in response to cytokine stimulation. The new approach presented in this study is generally applicable to any protein of interest for analyzing nucleocytoplasmic transport mechanisms in real time.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
There has been increasing evidence of basal nucleocytoplasmic shuttling of STAT3.
“As there has been increasing evidence of basal nucleocytoplasmic shuttling of STAT3 [6,41–43]”
From Selective STAT3-α or -β expression reveals spliceform-specific phosphorylation kinetics, nuclear retention and distinct gene expression outcomes (2012) · cites this paper as [42]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Principles of interleukin (IL)-6-type cytokine signalling and its regulation
HEINRICH, P. C., BEHRMANN, I., HAAN, S., HERMANNS, H. M., MÜLLER-NEWEN, G., & SCHAPER, F. (2003) · Biochemical Journal · doi:10.1042/bj20030407 · Cited by 3,272 · see citing works
Self-citationSTAT1 from the cell membrane to the DNA
Lillemeier, B. F., Köster, M., & Kerr, I. M. (2001) · The EMBO Journal · doi:10.1093/emboj/20.10.2508 · Cited by 88 · see citing works
Flagged first, then by citations
Research works printed but not linked: 5
- Takeda, K., Noguchi, K., Shi, W., Tanaka, T., Matsumoto, M., Yoshida, N., Kishimoto, T., and Akira, S. (1997) Proc. Natl. Acad. Sci. U. S. A. 94, 3801-3804
- Yoshida, K., Taga, T., Saito, M., Suematsu, S., Kumanogoh, A., Tanaka, T., Fujiwara, H., Hirata, M., Yamagami, T., Nakahata, T., Hirabayashi, T., Yoneda, Y., Tanaka, K., Wang, W. Z., Mori, C., Shiota, K., Yoshida, N., and Kishimoto, T. (1996) Proc. Natl. Acad. Sci. U. S. A. 93, 407-411
- McBride, K. M., and Reich, N. C. (2003) Science's STKE http://stke. sciencemag.org/cgi/content/full/sigtrans;2003/195/re13
- Lippincott-Schwartz, J., Snapp, E., and Kenworthy, A. (2001) Nat. Rev. Mol. Cell. Biol. 2, 444 -456
- Gauldie, J., Richards, C., Harnish, D., Lansdorp, P., and Baumann, H. (1987) Proc. Natl. Acad. Sci. U. S. A. 84, 7251-7255
Cited by
Sorted by citations
The potential and controversy of targeting STAT family members in cancer
Verhoeven, Y., Tilborghs, S., Jacobs, J., De Waele, J., Quatannens, D., Deben, C., Prenen, H., Pauwels, P., Trinh, X. B., Wouters, A., et al. (2020) · Seminars in Cancer Biology · doi:10.1016/j.semcancer.2019.10.002 · Cited by 409 · see citing works
Ng, I. H. W., Ng, D. C. H., Jans, D. A., & Bogoyevitch, M. A. (2012) · Biochemical Journal · doi:10.1042/bj20120941 · Cited by 63 · see citing works
Martincuks, A., Andryka, K., Küster, A., Schmitz-Van de Leur, H., Komorowski, M., & Müller-Newen, G. (2017) · Cellular Signalling · doi:10.1016/j.cellsig.2017.01.006 · Cited by 45 · see citing works
Self-citation
- Authors
- Pranada, A. L., Metz, S., Herrmann, A., Heinrich, P. C., & Müller-Newen, G.
- Affiliations
- Universitätsklinikum Aachen (DE)
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 2004-04
- Volume
- 279
- Issue
- 15
- Pages
- 15114-15123
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2011906109
- Cited by
- 140
- References
- 2
- Citations by year
- 2026: 3 · 2025: 1 · 2024: 3 · 2023: 3 · 2022: 2 · 2021: 8 · 2020: 4 · 2019: 8 · 2018: 4 · 2017: 6 · 2016: 8 · 2015: 4 · 2014: 5 · 2013: 7 · 2012: 8
- FWCI
- 5.78
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Journal of Biological Chemistry (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RWTH Publications (RWTH Aachen) (submittedVersion)
- Funders
- Deutsche Forschungsgemeinschaft (SFB 542)
- Universität Wien
- Topics
- Health Sciences › Medicine › Oncology › Cytokine Signaling Pathways and Interactions
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Biophysics › Advanced Fluorescence Microscopy Techniques
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › NF-κB Signaling Pathways
- Keywords
- STAT3, nucleocytoplasmic shuttling, tyrosine 705 phosphorylation, nuclear export, IL-6 signaling, cytokine induction
- MeSH terms
- Animals, Cell Line, Cell Nucleus, Cloning, Molecular, Cytoplasm, Cytosol, DNA-Binding Proteins, Electrophoresis, Polyacrylamide Gel, Humans, Lasers, Luminescent Proteins, Microscopy, Fluorescence, Mutation, Phosphorylation, Precipitin Tests, Protein Binding, Time Factors, Transfection, Tyrosine, Blotting, Western, Signal Transduction, Trans-Activators, Interleukin-6, Cytokines, Gene Deletion, Protein Structure, Tertiary, Genes, Reporter, Microscopy, Confocal, COS Cells, Protein Transport, Green Fluorescent Proteins, STAT3 Transcription Factor
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
