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Protein arginine methyltransferase 5 is a key regulator of the MYCN oncoprotein in neuroblastoma cells.

Park JH, Szemes M, Vieira GC, Melegh Z, Malik S, Heesom KJ, Von Wallwitz-Freitas L, Greenhough A, Brown KW, Zheng YG, Catchpoole D, Deery MJ, Malik K - Mol Oncol (2014)

Bottom Line: PRMT5 knockdown in MYCN-overexpressing cells, including the SHEP-21N cell-line with inducible MYCN expression leads to a dramatic decrease in MYCN protein and MYCN-associated cell-death in SHEP-21N cells.By using liquid chromatography - tandem mass spectrometry (LC-MS/MS) analysis of immunoprecipitated MYCN protein, we identified several potential sites of arginine dimethylation on the MYCN protein.Together our studies implicate PRMT5 in a novel mode of MYCN post-translational regulation and suggest PRMT5 plays a major role in NB tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Cancer Epigenetics Laboratory University of Bristol, Bristol BS8 1TD, UK.

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Physical interaction between endogenous PRMT5 and MYCN. (A) Co-immunoprecipitation (IP) of MYCN with anti-PRMT5 antibody (left) and the reciprocal co-immunoprecipitation (right) in NGP cells. (B) Co-immunoprecipitations as above in SK-N-BE(2)C cells.
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fig7: Physical interaction between endogenous PRMT5 and MYCN. (A) Co-immunoprecipitation (IP) of MYCN with anti-PRMT5 antibody (left) and the reciprocal co-immunoprecipitation (right) in NGP cells. (B) Co-immunoprecipitations as above in SK-N-BE(2)C cells.

Mentions: We next sought to establish whether endogenous PRMT5 physically associated with MYCN complexes in NGP and SK-N-BE(2)C cells, as such an interaction may serve to stabilise MYCN and provide initial evidence that MYCN might be a substrate for the arginine methyltransferase activity of PRMT5. As shown in Figure 7A, MYCN was easily detected in PRMT5 complexes immunoprecipitated from NGP extracts and PRMT5 was also confirmed in the reciprocal MYCN immunoprecipitation. This robust interaction was also apparent in SK-N-BE(2)C cells (Figure 7B), and is consistent with the notion that PRMT5 is capable of stabilizing MYCN at the protein level.


Protein arginine methyltransferase 5 is a key regulator of the MYCN oncoprotein in neuroblastoma cells.

Park JH, Szemes M, Vieira GC, Melegh Z, Malik S, Heesom KJ, Von Wallwitz-Freitas L, Greenhough A, Brown KW, Zheng YG, Catchpoole D, Deery MJ, Malik K - Mol Oncol (2014)

Physical interaction between endogenous PRMT5 and MYCN. (A) Co-immunoprecipitation (IP) of MYCN with anti-PRMT5 antibody (left) and the reciprocal co-immunoprecipitation (right) in NGP cells. (B) Co-immunoprecipitations as above in SK-N-BE(2)C cells.
© Copyright Policy - CC BY
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4359099&req=5

fig7: Physical interaction between endogenous PRMT5 and MYCN. (A) Co-immunoprecipitation (IP) of MYCN with anti-PRMT5 antibody (left) and the reciprocal co-immunoprecipitation (right) in NGP cells. (B) Co-immunoprecipitations as above in SK-N-BE(2)C cells.
Mentions: We next sought to establish whether endogenous PRMT5 physically associated with MYCN complexes in NGP and SK-N-BE(2)C cells, as such an interaction may serve to stabilise MYCN and provide initial evidence that MYCN might be a substrate for the arginine methyltransferase activity of PRMT5. As shown in Figure 7A, MYCN was easily detected in PRMT5 complexes immunoprecipitated from NGP extracts and PRMT5 was also confirmed in the reciprocal MYCN immunoprecipitation. This robust interaction was also apparent in SK-N-BE(2)C cells (Figure 7B), and is consistent with the notion that PRMT5 is capable of stabilizing MYCN at the protein level.

Bottom Line: PRMT5 knockdown in MYCN-overexpressing cells, including the SHEP-21N cell-line with inducible MYCN expression leads to a dramatic decrease in MYCN protein and MYCN-associated cell-death in SHEP-21N cells.By using liquid chromatography - tandem mass spectrometry (LC-MS/MS) analysis of immunoprecipitated MYCN protein, we identified several potential sites of arginine dimethylation on the MYCN protein.Together our studies implicate PRMT5 in a novel mode of MYCN post-translational regulation and suggest PRMT5 plays a major role in NB tumorigenesis.

View Article: PubMed Central - PubMed

Affiliation: Cancer Epigenetics Laboratory University of Bristol, Bristol BS8 1TD, UK.

Show MeSH
Related in: MedlinePlus