Abstract
Similar to ubiquitin, regulatory roles for NEDD8 (neural precursor cell-expressed developmentally down-regulated 8) are being clarified during cell growth, signal transduction, immune response, and development. However, NEDD8 targets and their functional alterations are not well known. Regulator of calcineurin 1 (RCAN1/DSCR1P1) is located near the Down syndrome critical region on the distal part of chromosome 21, and its gene product is an endogenous inhibitor of calcineurin signaling. RCAN1 is modified by ubiquitin and consequently undergoes proteasomal degradation. Here we report that NEDD8 is conjugated to RCAN1 (RCAN1-1S) via three lysine residues, K96, K104, and K107. Neddylation enhances RCAN1 protein stability without affecting its cellular location. In addition, we found that neddylation significantly inhibits proteasomal degradation of RCAN1, which may underlie the ability of NEDD8 to enhance RCAN1 stability. Furthermore, neddylation increases RCAN1 binding to calcineurin, which potentiates its inhibitory activity toward downstream NFAT signaling. The present study provides a new regulatory mechanism of RCAN1 function and highlights an important role for diverse RCAN1-involved cellular physiology.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Binding Sites
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COS Cells
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Calcineurin / metabolism
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Calcineurin Inhibitors*
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Cell Nucleus / metabolism
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Chlorocebus aethiops
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Cytosol / metabolism
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DNA-Binding Proteins
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HEK293 Cells
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Humans
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Intracellular Signaling Peptides and Proteins / chemistry*
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Intracellular Signaling Peptides and Proteins / metabolism*
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Mice
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Muscle Proteins / chemistry*
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Muscle Proteins / metabolism*
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NEDD8 Protein
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NFATC Transcription Factors / metabolism
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Protein Binding
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Protein Stability
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Signal Transduction
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Ubiquitins / metabolism*
Substances
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Calcineurin Inhibitors
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DNA-Binding Proteins
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Intracellular Signaling Peptides and Proteins
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Muscle Proteins
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NEDD8 Protein
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NEDD8 protein, human
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NFATC Transcription Factors
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RCAN1 protein, human
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Ubiquitins
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Calcineurin
Grants and funding
This study was supported by grants from the Korea Healthcare Technology R&D Project (A092004 and A111653 to K.C.C.) funded by the Ministry for Health, Welfare & Family Affairs, Republic of Korea. This work was also partially supported by grants from the Brain Research Center of the 21st Century Frontier Research Program Technology (2009K-001251 to K.C.C.), the National Research Foundation of Korea (NRF; 2010-0018916 to K.C.C.), and the Basic Science Research Program through NRF (2012-0000810 to K.C.C.) funded by the Ministry of Education, Science and Technology (MEST), Republic of Korea. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.