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Functional importance of RASSF1A microtubule localization and polymorphisms

Oncogene volume 29, pages 5729–5740 (2010)Cite this article

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Abstract

Ras association domain family protein 1A (RASSF1A) is one of the more heavily methylated genes in human cancers. In addition to promoter-specific methylation, RASSF1A polymorphisms have been identified in cancer patients. RASSF1A is a tumor suppressor protein involved in death receptor-dependent apoptosis and it is localized to microtubules. Currently, the biological importance of RASSF1A microtubule localization and the functional consequences of RASSF1A polymorphisms is not understood. In this study, we have investigated both RASSF1A microtubule association and polymorphisms. Loss of RASSF1A microtubule association resulted in the nuclear appearance of RASSF1A and the loss of association with α-, γ- and β-tubulin. Moreover, the loss of microtubule localization of RASSF1A resulted in enhanced tumor-promoting potential, as determined by a xenograft transplantation model in nude mice. It is surprising that, several RASSF1A polymorphisms also lost the ability to associate with α-, γ- and β-tubulin and lost the ability to prevent tumor formation in a xenograft nude mouse model when compared with wild-type RASSF1A. Our results demonstrate a role for RASSF1A microtubule localization in eliciting its tumor suppressor function. In addition, some RASSF1A polymorphisms lack the tumor suppressor function of RASSF1A and, if present in patients, may be tumorigenic.

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Accession codes

Accessions

GenBank/EMBL/DDBJ

Abbreviations

RASSF:

Ras association domain family

TNFα:

tumor necrosis factor α

TNF-R1:

TNFα receptor R1

MOAP-1:

modulator of apoptosis 1

TRAIL:

TNFα apoptosis-inducing ligand

TRAIL-R1:

TRAIL receptor 1

PARP:

poly (ADP-ribose) polymerase

NFκB:

nuclear factor κB

APC1:

adenomatous polyposis coli

GSK-3β:

glycogen synthase kinase 3β

GFP:

green fluorescent protein

GST:

glutathione S-transferase

HA:

Hematagluttin

WT:

wild type

SDS–PAGE:

sodium dodecyl sulphate gel electrophoresis

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Acknowledgements

We thank Dr Haya Abu Ghazaleh for her helpful discussions, Adrienne DeCorby-Baksh for her excellent technical assistance with our confocal imaging, Dr Victor Yu and Naiyang Fu for the MOAP-1 expression construct. This work was supported by grants from CIHR (MOP-79494) (MEK), the Women and Children's Health Research Institute (WCO14) (MEK), the Stollery Children's Hospital Foundation Donation Grant (PD946) (SB and MEK), Alberta Heritage Foundation for Medical Research (G220170170) (SB), and Canadian Foundation for Innovation/Alberta Small Equipment Grants Program (#13118).

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  1. Department of Pediatrics, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, Alberta, Canada

    M El-Kalla, C Onyskiw & S Baksh

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  1. M El-Kalla
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  2. C Onyskiw
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Correspondence to S Baksh.

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El-Kalla, M., Onyskiw, C. & Baksh, S. Functional importance of RASSF1A microtubule localization and polymorphisms. Oncogene 29, 5729–5740 (2010). https://doi.org/10.1038/onc.2010.316

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