N-terminal and C-terminal domains of calmodulin mediate FADD and TRADD interaction

Giuliana Papoff; Nadia Trivieri; Sonia Marsilio; Roberta Crielesi; Cristiana Lalli; Loriana Castellani; Edward M Balog; Giovina Ruberti

doi:10.1371/journal.pone.0116251

N-terminal and C-terminal domains of calmodulin mediate FADD and TRADD interaction

PLoS One. 2015 Feb 2;10(2):e0116251. doi: 10.1371/journal.pone.0116251. eCollection 2015.

Authors

Giuliana Papoff¹, Nadia Trivieri¹, Sonia Marsilio¹, Roberta Crielesi¹, Cristiana Lalli¹, Loriana Castellani², Edward M Balog³, Giovina Ruberti¹

Affiliations

¹ National Research Council, Institute of Cell Biology and Neurobiology, Campus Adriano Buzzati-Traverso, Monterotondo, Rome, Italy.
² National Research Council, Institute of Cell Biology and Neurobiology, Campus Adriano Buzzati-Traverso, Monterotondo, Rome, Italy; Department of Human Sciences, Society and Health, University of Cassino, Cassino, Italy.
³ School of Applied Physiology, Georgia Institute of Technology, Atlanta, Georgia, United States of America.

Abstract

FADD (Fas-associated death domain) and TRADD (Tumor Necrosis Factor Receptor 1-associated death domain) proteins are important regulators of cell fate in mammalian cells. They are both involved in death receptors mediated signaling pathways and have been linked to the Toll-like receptor family and innate immunity. Here we identify and characterize by database search analysis, mutagenesis and calmodulin (CaM) pull-down assays a calcium-dependent CaM binding site in the α-helices 1-2 of TRADD death domain. We also show that oxidation of CaM methionines drastically reduces CaM affinity for FADD and TRADD suggesting that oxidation might regulate CaM-FADD and CaM-TRADD interactions. Finally, using Met-to-Leu CaM mutants and binding assays we show that both the N- and C-terminal domains of CaM are important for binding.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Amino Acid Sequence
Binding Sites
Calcium / metabolism
Calmodulin / chemistry*
Calmodulin / metabolism*
Cell Line
Fas-Associated Death Domain Protein / metabolism*
Humans
Methionine / metabolism
Methionine Sulfoxide Reductases / pharmacology
Molecular Sequence Data
Mutation
Oxidation-Reduction
Protein Binding / drug effects
Protein Structure, Tertiary
TNF Receptor-Associated Death Domain Protein / chemistry
TNF Receptor-Associated Death Domain Protein / genetics
TNF Receptor-Associated Death Domain Protein / metabolism*

Substances

Calmodulin
FADD protein, human
Fas-Associated Death Domain Protein
TNF Receptor-Associated Death Domain Protein
Methionine
Methionine Sulfoxide Reductases
methionine sulfoxide reductase
Calcium

Grants and funding

This work was supported by grants (to G. R.) from the National Research Council: i) Project RTL “FADD and TRADD non apoptotic functions”; ii) Project “FaReBio di Qualità” financed by the Italian Ministry of Economy and Finance. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.