Interaction of the growth and tumour suppressor NORE1A with microtubules is not required for its growth-suppressive function.

BMC Research Notes
Anna MoshnikovaAndrei Khokhlatchev

Abstract

The NORE1 protein was identified in a yeast two-hybrid screen as a Ras effector that binds Ras protein in a GTP-dependent manner. NORE1A is a growth and tumour suppressor that is inactivated in a variety of cancers. In transformed human cells, both full-length NORE1A protein and its effector domain alone (amino acids 191-363) are localized to microtubules and centrosomes. However, the mechanism by which NORE1A associates with these cytoskeletal elements is not known; furthermore, whether centrosomally-associated or microtubule-associated NORE1A suppresses tumour cell growth has not been yet established. We have shown that purified NORE1A fails to bind to microtubules in vitro suggesting that other protein(s) mediate NORE1A-microtubule association. Using mass-spectrometry, we identified the Microtubule-Associated Protein 1B (MAP1B) and its homologue C19ORF5 as NORE1A interaction partners. Suppression of C19ORF5 expression by RNA interference (RNAi) and immunodepletion of C19ORF5 protein from cell extracts showed that binding of NORE1A to microtubules is not dependent on C19ORF5. Conversely, RNAi suppression of MAP1B revealed that MAP1B is required for association of NORE1A with microtubules. RNAi-mediated depletion of C19ORF5 or...Continue Reading

Citations

Aug 21, 2013·Advances in Biological Regulation·Jia Jia Chan, Matilda Katan
Feb 25, 2014·Tumour Biology : the Journal of the International Society for Oncodevelopmental Biology and Medicine·Li-Li LiuChris Zhiyi Zhang
May 4, 2010·Lung Cancer : Journal of the International Association for the Study of Lung Cancer·Kazuya ShinmuraHaruhiko Sugimura
Oct 30, 2013·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Takeshi NakamuraNaoyuki Wada
Sep 17, 2014·The Journal of Biological Chemistry·Howard DonningerGeoffrey J Clark
Mar 16, 2017·Protoplasma·Tomáš Mazel
Jun 15, 2014·Journal of Cell Science·Rui YangAndreas W Püschel

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Methods Mentioned

BETA
coimmunoprecipitation
immunodepletion
flow cytometry
Transfection
Immunoprecipitation
Protein
Spin
Assay

Software Mentioned

Adobe Photoshop
ModFIT
IPLab

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