Ras Transformation Overrides a Proliferation Defect Induced by Tpm3.1 Knockout.

Abstract

Extensive re-organisation of the actin cytoskeleton and changes in the expression of its binding proteins is a characteristic feature of cancer cells. Previously we have shown that the tropomyosin isoform Tpm3.1, an integral component of the actin cytoskeleton in tumor cells, is required for tumor cell survival. Our objective was to determine whether cancer cells devoid of Tpm3.1 would evade the tumorgenic effects induced by H-Ras transformation. The tropomyosin isoform (Tpm) expression profile of a range of cancer cell lines (21) demonstrates that Tpm3.1 is one of the most broadly expressed Tpm isoform. Consequently, the contribution of Tpm3.1 to the transformation process was functionally evaluated. Primary embryonic fibroblasts isolated from wild type (WT) and Tpm3.1 knockout (KO) mice were transduced with retroviral vectors expressing SV40 large T antigen and an oncogenic allele of the H-Ras gene, H-RasV12, to generate immortalized and transformed WT and KO MEFs respectively. We show that Tpm3.1 is required for growth factor-independent proliferation in the SV40 large T antigen immortalized MEFs, but this requirement is overcome by H-Ras transformation. Consistent with those findings, we found that Tpm3.1 was not required for anchorage independent growth or growth of H-Ras-driven tumors in a mouse model. Finally, we show that pERK and Importin 7 protein interactions are significantly decreased in the SV40 large T antigen immortalized KO MEFs but not in the H-Ras transformed KO cells, relative to control MEFs. The data demonstrate that H-Ras transformation overrides a requirement for Tpm3.1 in growth factor-independent proliferation of immortalized MEFs. We propose that in the SV40 large T antigen immortalized MEFs, Tpm3.1 is partly responsible for the efficient interaction between pERK and Imp7 resulting in cell proliferation, but this is overidden by Ras transformation.

• Authors: Jason D Coombes, Galina Schevzov, Chin-Yi Kan, Carlotta Petti, Michelle F Maritz, Shane Whittaker, Karen L Mackenzie, Peter W Gunning
• Journal: Cellular & molecular biology letters (Cell Mol Biol Lett) Vol. 20 Issue 4 Pg. 626-46 (Dec 2015) ISSN: 1689-1392 [Electronic] England
• PMID: 26274783 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antigens, Polyomavirus Transforming
• Karyopherins
• Protein Isoforms
• TPM3 protein, human
• Tpm3 protein, mouse
• Tropomyosin
• Ipo7 protein, mouse
• Extracellular Signal-Regulated MAP Kinases

Topics

• Animals
• Antigens, Polyomavirus Transforming (genetics)
• Cell Line, Tumor
• Cell Proliferation
• Cell Transformation, Neoplastic (genetics)
• Cells, Cultured
• Extracellular Signal-Regulated MAP Kinases (metabolism)
• Female
• Fibroblasts (pathology)
• Gene Expression Regulation, Neoplastic
• Genes, ras (genetics)
• Humans
• Karyopherins (genetics, metabolism)
• Mice, Inbred BALB C
• Mice, Inbred C57BL
• Mice, Knockout
• Protein Isoforms (genetics, metabolism)
• Tropomyosin (genetics, metabolism)