Binding to Intracellular Targets of the Metastasis-Inducing Protein, S100A4 (p9Ka)

doi:10.1006/bbrc.2001.5517

Biochemical and Biophysical Research Communications

Volume 286, Issue 5, 7 September 2001, Pages 1212-1217

https://doi.org/10.1006/bbrc.2001.5517 Get rights and content

Abstract

Experimentally elevated levels of S100A4 induce a metastatic phenotype in benign mammary tumour cells in vivo. In humans, the presence of S100A4 in breast cancer cells correlates strongly with reduced patient survival. Potential interacting binding partners for S100A4 have now been examined using an optical biosensor. There was significant interaction of S100A4 with non-muscle myosin and p53, but not with actin, tropomyosin or tubulin. The results suggest that myosin and p53 are likely to be intracellular targets of S100A4. S100A4 had a greater affinity for wild-type or mutant arg-175-his p53 than for non-muscle myosin. The results suggest that S100A4 might induce metastasis by influencing the function of p53 as well as through its interaction with myosin and that any mechanism is independent of the mutational status of p53.

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Cited by (77)

S100A4/Nonmuscle Myosin IIA/p53 Axis Contributes to Aggressive Features in Ovarian High-Grade Serous Carcinoma
2020, American Journal of Pathology
S100A4 is a small calcium-binding protein that exerts its biological functions by interacting with nonmuscle myosin IIA (NMIIA) and p53. Although S100A4 promotes metastasis in several tumors, little is known about its involvement in the progression of ovarian high-grade serous carcinomas (HGSCs). Herein, we focused on functional roles of the S100A4/NMIIA/p53 axis in these tumors. In HGSC cell lines harboring mutant p53, knockdown (KD) of S100A4 reduced the expression of several epithelial-mesenchymal transition/cancer stem cell markers and the ALDH1^high population, consistent with an inhibition of stemness features. S100A4-KD also increased apoptosis, decreased cell proliferation, and accelerated cell mobility. This was accompanied by increased Snail expression, which, in turn, was likely due to loss of p53 function. In contrast, specific inhibition of NMIIA by blebbistatin induced phenotypes that—with the exception of cell proliferation and mobility—were opposite to those observed in S100A4-KD cells. In clinical samples, cytoplasmic and/or nuclear interactions between S100A4, NMIIA, and mutant p53 were observed. In addition, high expression of S100A4, but not NMIIA or p53, was a significant and independent unfavorable prognostic factor in HGSC patients. These findings suggest that, via its interaction with NMIIA and p53, overexpressed S100A4 may induce epithelial-mesenchymal transition/cancer stem cell properties in HGSC and elicit several other tumor-associated phenotypes.
Pentamidine inhibit S100A4 - p53 interaction and decreases cell proliferation activity
2020, Archives of Biochemistry and Biophysics
Metastasis-associated S100A4 protein is a small calcium-binding protein typically overexpressed in several tumor forms, and it is widely accepted that S100A4 plays a significant role in the metastasis of cancer. Tumor suppressor p53 is one of the S100A4's main targets. Previous reports show that through p53, S100A4 regulates collagen expression and cell proliferation. When S100A4 interacts with p53, the S100A4 destabilizes wild type p53. In the current study, based on ¹H–¹⁵N HSQC NMR experiments and HADDOCK results, S100A4 interacts with the intrinsically unstructured transactivation domain (TAD) of the protein p53 and the pentamidine molecules in the presence of calcium ions. Our results suggest that the p53 TAD and pentamidine molecules share similar binding sites on the S100A4 protein. This observation indicates that a competitive binding mechanism can interfere with the binding of S100A4-p53 and increase the level of p53. Also, we compare different aspects of p53 activity in the WST-1 test using MCF 7 cells. We found that the presence of a pentamidine molecule results in higher p53 activity, which is also reflected in less cell proliferation. Collectively, our results indicate that disrupting the S100A4-p53 interaction would prevent cancer progression, and thus S100A4-p53 inhibitors provide a new avenue for cancer therapy.
Role of S100 proteins in health and disease
2020, Biochimica et Biophysica Acta - Molecular Cell Research
The S100 family of proteins contains 25 known members that share a high degree of sequence and structural similarity. However, only a limited number of family members have been characterized in depth, and the roles of other members are likely undervalued. Their importance should not be underestimated however, as S100 family members function to regulate a diverse array of cellular processes including proliferation, differentiation, inflammation, migration and/or invasion, apoptosis, Ca²⁺ homeostasis, and energy metabolism. Here we detail S100 target protein interactions that underpin the mechanistic basis to their function, and discuss potential intervention strategies targeting S100 proteins in both preclinical and clinical situations.
S100A4 regulates the Src-tyrosine kinase dependent differentiation of Th17 cells in rheumatoid arthritis
2014, Biochimica et Biophysica Acta - Molecular Basis of Disease
Objectives: To evaluate the role of S100A4, a calcium-binding regulator of nonmuscle myosin assembly, for T-cell responses in rheumatoid arthritis. Methods: Arthritis was induced in the methylated bovine serum albumin (mBSA)-immunized mice lacking the entire S100A4 protein (S100A4KO) and in wild-type counterparts treated with short hairpin ribonucleic acid (shRNA)-lentiviral constructs targeting S100A4 (S100A4-shRNA). The severity of arthritis was evaluated morphologically. T-cell subsets were characterized by the expression of master transcription factors, and functionally by proliferation activity and cytokine production. The activity of the Scr-kinases Fyn and Lck was assessed by the autophosphorylation of C-terminal thyrosine and by the phosphorylation of the CD5 cytodomain. The interaction between S100A4 and the CD5 cytodomain was analysed by nuclear magnetic resonance spectrophotometry. Results: S100A4-deficient mice (S100A4KO and S100A4-shRNA) had significantly alleviated morphological signs of arthritis and joint damage. Leukocyte infiltrates in the arthritic joints of S100A4-deficient mice accumulated Foxp3⁺ Treg cells, while the number of RORγt⁺ and (pTyr705)STAT3⁺ cells was reduced. S100A4-deficient mice had a limited formation of Th17-cells with low retinoic acid orphan receptor gamma t (RORγt) mRNA and IL17 production in T-cell cultures. S100A4-deficient mice had a low expression and activity of T-cell receptor (TCR) inhibitor CD5 and low (pTyr705)STAT3 (signal transducer and activator of transcription 3), which led to increased (pTyr352)ZAP-70 (theta-chain associated protein kinase of 70 kDa), lymphocyte proliferation and production of IL2. In vitro experiments showed that S100A4 directly binds Lck and Fyn and reciprocally regulates their kinase activity towards the CD5 cytodomain. Spectrometry demonstrates an interaction between the CD5 cytodomain and EF2-binding sites of S100A4. Conclusion: The present study demonstrates that S100A4 plays an important part in the pathogenesis of arthritis. It controls CD5-dependent differentiation of Th17 cells by regulating the activity of the Src-family kinases Lck and Fyn.
S100A6 competes with the TAZ2 domain of p300 for binding to p53 and attenuates p53 acetylation
2013, Journal of Molecular Biology
Citation Excerpt :
In the context of the potential involvement of S100 proteins in cancer, it is important to note that some of them, including S100A6, are able to bind p53, a protein crucial for the control of cell proliferation and apoptosis [9]. The effect of the S100–p53 interaction on p53 transcriptional activity was found to be positive in the case of S100A2 and S100A6 [10,11] while the effect of S100A4 and S100B proteins differed depending on cells and conditions applied [12–15]. Biophysical studies showed differences in the interaction sites and p53 binding affinities between individual S100 proteins, which could partly explain their disparate effects on p53 activity.
S100A6 is a calcium binding protein that, like some other members of the S100 protein family, is able to bind p53. This interaction may be physiologically relevant considering the numerous connotations of S100 proteins and of S100A6, in particular, with cancer and metastasis. In this work, we show that the interaction with S100A6 is limited to unmodified or phosphorylated p53 and is inhibited by p53 acetylation. Using in vitro acetylation assay, we show that the presence of S100A6 attenuates p53 acetylation by p300. Furthermore, using ELISA, we show that S100A6 and the TAZ2 domain of p300 bind p53 with similar affinities and that S100A6 effectively competes with TAZ2 for binding to p53. Our results add another element to the complicated scheme of p53 activation.
S100P dissociates myosin IIA filaments and focal adhesion sites to reduce cell adhesion and enhance cell migration
2012, Journal of Biological Chemistry
S100 proteins promote cancer cell migration and metastasis. To investigate their roles in the process of migration we have constructed inducible systems for S100P in rat mammary and human HeLa cells that show a linear relationship between its intracellular levels and cell migration. S100P, like S100A4, differentially interacts with the isoforms of nonmuscle myosin II (NMIIA, K_d = 0.5 μm; IIB, K_d = 8 μm; IIC, K_d = 1.0 μm). Accordingly, S100P dissociates NMIIA and IIC filaments but not IIB in vitro. NMIIA knockdown increases migration in non-induced cells and there is no further increase upon induction of S100P, whereas NMIIB knockdown reduces cell migration whether or not S100P is induced. NMIIC knockdown does not affect S100P-enhanced cell migration. Further study shows that NMIIA physically interacts with S100P in living cells. In the cytoplasm, S100P occurs in discrete nodules along NMIIA-containing filaments. Induction of S100P causes more peripheral distribution of NMIIA filaments. This change is paralleled by a significant drop in vinculin-containing, actin-terminating focal adhesion sites (FAS) per cell. The induction of S100P, consequently, causes significant reduction in cellular adhesion. Addition of a focal adhesion kinase (FAK) inhibitor reduces disassembly of FAS and thereby suppresses S100P-enhanced cell migration. In conclusion, this work has demonstrated a mechanism whereby the S100P-induced dissociation of NMIIA filaments leads to a weakening of FAS, reduced cell adhesion, and enhanced cell migration, the first major step in the metastatic cascade.

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¹: Present address: Department of Medicine, University of Cambridge, Cambridge, UK.

²: To whom correspondence should be addressed at Life Sciences Building, School of Biological Sciences, University of Liverpool, P.O. Box 147, Liverpool L69 7ZB, UK. Fax: 44 151 794 4349. E-mail: [email protected].

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Regular ArticleBinding to Intracellular Targets of the Metastasis-Inducing Protein, S100A4 (p9Ka)

Abstract

Molecular cloning and sequence of the gene for p9Ka, a cultured myoepithelial cell protein with strong homology to S-100, a calcium-binding protein

J. Mol. Biol.

Induction of the metastatic phenotype by transfection of a benign rat mammary epithelial cell line with the gene for p9Ka, a rat calcium-binding protein but not with the oncogene EJ ras-1

Oncogene

Human S100A4 (p9Ka) induces the metastatic phenotype upon benign tumour cells

Oncogene

Expression of the calcium-binding protein S100A4 (p9Ka) in MMTV-neu transgenic mice induces metastasis of mammary tumours

Oncogene

Metastasis of mammary carcinomas in GRS/A hybrid mice transgenic for the mts1 gene

Oncogene

Prognostic significance of the metastasis-inducing protein S100A4 (p9Ka) in human breast cancer

Cancer Res.

Comparison of the metastasis-inducing protein S100A4 (p9Ka) with other prognostic markers in human breast cancer

Int. J. Cancer (Pred. Oncol.)

Calvasculin as a factor affecting the microfilament assemblies in rat fibroblasts transfected by src gene

FEBS Lett.

Binding of pEL98 protein, an S100-related calcium-binding protein, to non-muscle tropomyosin

J. Cell Biol.

Interaction of metastasis associated mts1 protein with non-muscle myosin

Oncogene

Effect of Mts1 on the structure and activity of nonmuscle myosin II

Biochemistry

Non-muscle myosin heavy chain as a possible target for protein encoded by metastasis-related mts-1 gene

J. Biol. Chem.

Metastasis-associated Mts1 (S100A4) protein modulates protein kinase C phosphorylation of the heavy chain of nonmuscle myosin

J. Biol. Chem.

Metastasis associated MTS1 and NM23 genes affect tubulin polymerisation in B16 melanomas: A possible mechanism of their regulation of metastatic behaviour of tumours

Anticancer Res.

Metastasis-associated mts1 gene expression correlates with increased p53 detection in the B16 melanoma

DNA and Cell Biol.

Tumor suppressor p53 protein is a new target for the metastasis-associated Mts1/S10A4 protein: Functional consequences of their interaction

J. Biol. Chem.

Regular Article
Binding to Intracellular Targets of the Metastasis-Inducing Protein, S100A4 (p9Ka)