Extracellular vesicles (EVs) are produced by most prokaryotic and eukaryotic cells; tumour cells, however, release much higher amounts of EVs, which contain cancer-specific proteins and RNAs. Molecules carried by EVs are captured by surrounding cells, which then undergo profound phenotypic modifications. G26/24 oligodendroglioma cells release, for example, EVs containing FasL and TRAIL, which induce apoptosis in rat cortical neurons and astrocytes in culture. By metabolic labelling of cells, EV-mediated horizontal transfer of radioactive proteins was clearly demonstrated. Among the proteins present in EVs produced by oligodendroglioma cells, extracellular matrix remodelling proteases, and the linker histone H1°, a differentiation-specific histone, were identified. We also found that A375 melanoma cells release EVs which, like the once produced by oligodendroglioma cells, contain H1°. Interestingly, H1° histone sorted to vesicles has a molecular mass higher than expected, and is probably sumoylated. More recently, by a T1 RNase-protection assay, done by mixing an in-vitro transcribed, H1°- encoding RNA, and EVs, three main RNA-protein complexes were evidenced, the most abundant of which had an apparent molecular mass of about 65 kDa. We then synthesized a biotinylated H1° RNA, with the aim to fish, by affinity chromatography, the evidenced proteins. The RNA-bound fraction was finally analysed by mass spectrometry. The most abundant protein identified was the myelin expression factor-2 (MYEF2), which has indeed a molecular mass of about 60 kDa. The presence of MYEF2 in EVs released from A375 melanoma cells was finally confirmed by Western blot analysis.
Schiera, G., Di Liegro, C., Puleo, V., Colletta, O., & Di Liegro, I. (2015). Melanoma cells release extracellular vesicles which contain RNA-binding proteins able to bind the mRNA encoding histone H1°. In ABSTRACT BOOK (pp.28-28).
|Autori:||Schiera, G.; Di Liegro, C.; Puleo, V.; Colletta, O.; Di Liegro, I.|
|Titolo:||Melanoma cells release extracellular vesicles which contain RNA-binding proteins able to bind the mRNA encoding histone H1°|
|Settore Scientifico Disciplinare:||Settore BIO/10 - Biochimica|
Settore BIO/06 - Anatomia Comparata E Citologia
|Nome del convegno:||ISEV Research seminar Extracellular vesicle-associated RNA: is there a purpose?|
|Luogo del convegno:||Utrecht|
|Anno del convegno:||dal 24-09-2015 al 25-09-2015|
|Data di pubblicazione:||2015|
|Numero di pagine:||1|
|Citazione:||Schiera, G., Di Liegro, C., Puleo, V., Colletta, O., & Di Liegro, I. (2015). Melanoma cells release extracellular vesicles which contain RNA-binding proteins able to bind the mRNA encoding histone H1°. In ABSTRACT BOOK (pp.28-28).|
|Tipologia:||0 - Proceedings (TIPOLOGIA NON ATTIVA)|
|Appare nelle tipologie:||0 - Proceedings (TIPOLOGIA NON ATTIVA)|
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