Dominant-negative function of the C-terminal fragments of NBR1 and SQSTM1 generated during enteroviral infection.

Abstract

Coxsackievirus infection induces an abnormal accumulation of ubiquitin aggregates that are generally believed to be noxious to the cells and have a key role in viral pathogenesis. Selective autophagy mediated by autophagy adaptor proteins, including sequestosome 1 (SQSTM1/p62) and neighbor of BRCA1 gene 1 protein (NBR1), are an important pathway for disposing of misfolded/ubiquitin conjugates. We have recently demonstrated that SQSTM1 is cleaved after coxsackievirus infection, resulting in the disruption of SQSTM1 function in selective autophagy. NBR1 is a functional homolog of SQSTM1. In this study, we propose to test whether NBR1 can compensate for the compromise of SQSTM1 after viral infection. Of interest, we found that NBR1 was also cleaved after coxsackievirus infection. This cleavage took place at two sites mediated by virus-encoded protease 2A(pro) and 3C(pro), respectively. In addition to the loss-of-function, we further investigated whether cleavage of SQSTM1/NBR1 leads to the generation of toxic gain-of-function mutants. We showed that the C-terminal fragments of SQSTM1 and NBR1 exhibited a dominant-negative effect against native SQSTM1/NBR1, probably by competing for LC3 and ubiquitin chain binding. Finally, we demonstrated a positive, mutual regulatory relationship between SQSTM1 and NBR1 during viral infection. We showed that knockdown of SQSTM1 resulted in reduced expression of NBR1, whereas overexpression of SQSTM1 led to increased level of NBR1, and vice versa, further excluding the possible compensation of NBR1 for the loss of SQSTM1. Taken together, the findings in this study suggest a novel mechanism through which coxsackievirus infection induces increased accumulation of ubiquitin conjugates and subsequent viral damage.

• Authors: J Shi, G Fung, P Piesik, J Zhang, H Luo
• Journal: Cell death and differentiation (Cell Death Differ) Vol. 21 Issue 9 Pg. 1432-41 (Sep 2014) ISSN: 1476-5403 [Electronic] England
• PMID: 24769734 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Adaptor Proteins, Signal Transducing
• Intracellular Signaling Peptides and Proteins
• NBR1 protein, human
• Proteins
• SQSTM1 protein, human
• Sequestosome-1 Protein

Topics

• Adaptor Proteins, Signal Transducing (biosynthesis, genetics, metabolism)
• Enterovirus Infections (metabolism)
• Genes, Dominant
• HeLa Cells
• Humans
• Intracellular Signaling Peptides and Proteins
• Proteins (genetics, metabolism)
• Sequestosome-1 Protein
• Tumor Cells, Cultured