The cytosolic sialidase Neu2 is degraded by autophagy during myoblast atrophy

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• 作者：Stefania Rossi ; Elena Stoppani ; Wim Martinet ; Andrea Bonetto ; Paola Costelli ; Roberta Giuliani ; Francesca Colombo ; Augusto Preti ; Sergio Marchesini ; Aless ; ro Fanzani
• 关键词：Sialidases ; C2C12 myoblasts ; Atrophy ; Autophagy ; Lysosomes ; Cathepsin
• 刊名：Biochimica et Biophysica Acta (BBA)/General Subjects
• 出版年：2009
• 出版时间：August 2009
• 年：2009
• 卷：1790
• 期：8
• 页码：817-828
• 全文大小：1833 K

文摘

Background

The sialidase Neu2 is a cytosolic enzyme which is fully expressed in mature muscle myofibers.

Methods

To investigate Neu2 expression during muscle atrophy, we employed an in vitro model consisting of terminally differentiated C2C12 myotubes exposed to different pro-atrophic stimuli that triggered catabolic pathways involved in proteasome activation or autophagy.

Results

Neu2 expression was unchanged in myotubes treated with TNF-alpha, a cytokine known to activate the proteasome. However, Neu2 transcript levels and enzymatic activity were downregulated in starved or dexamethasone-treated myotubes that showed proteosomal activation and several hallmarks of macroautophagy, such as formation of autophagosomes, the accumulation of LC3 dots and bulk degradation of long-lived proteins. Neu2 activity and protein levels were rescued upon cotreatment with the lysosomotropic agent NH₄Cl, the autophagy inhibitor 3-methyladenine or cathepsin inhibitors, as well as by insulin administration, but were unaffected upon pharmacological inhibition of the proteasome. Moreover, HA- or GST-Neu2 recombinant fusion proteins were rapidly degraded in vitro by purified cathepsin L and B. Overall, we may conclude that Neu2 is degraded by lysosomal enzymes in myotubes undergoing autophagy-mediated atrophy.

General significance

This study demonstrates that Neu2 enzyme degradation occurs in atrophic myotubes via macroautophagy and independently of proteasome activation.