Bruton's tyrosine kinase phosphorylates DDX41 and activates its binding of dsDNA and STING to initiate type 1 interferon response

Koon-Guan Lee; Susana Soo-Yeon Kim; Lin Kui; Dominic Chih-Cheng Voon; Marjorie Mauduit; Pradeep Bist; Xuezhi Bi; Natasha Ann Pereira; Chengcheng Liu; Bindu Sukumaran; Laurent Rénia; Yoshiaki Ito; Kong-Peng Lam

doi:10.1016/j.celrep.2015.01.039

Bruton's tyrosine kinase phosphorylates DDX41 and activates its binding of dsDNA and STING to initiate type 1 interferon response

Cell Rep. 2015 Feb 24;10(7):1055-65. doi: 10.1016/j.celrep.2015.01.039. Epub 2015 Feb 19.

Affiliations

¹ Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore 138668, Singapore.
² Cancer Biology Program, Cancer Science Institute, National University of Singapore, Singapore 117599, Singapore.
³ Singapore Immunology Network, Agency for Science, Technology and Research, Singapore 138648, Singapore.
⁴ Emerging Infectious Diseases Program, DUKE-NUS Graduate Medical School, Singapore 169857, Singapore.
⁵ Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore 138668, Singapore; Departments of Microbiology, Physiology, and Paediatrics, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117574, Singapore. Electronic address: [email protected].

PMID: 25704810
DOI: 10.1016/j.celrep.2015.01.039

Abstract

The innate immune system senses cytosolic dsDNA and bacterial cyclic dinucleotides and initiates signaling via the adaptor STING to induce type 1 interferon (IFN) response. We demonstrate here that BTK-deficient cells have impaired IFN-β production and TBK1/IRF3 activation when stimulated with agonists or infected with pathogens that activate STING signaling. BTK interacts with STING and DDX41 helicase. The kinase and SH3/SH2 interaction domains of BTK bind, respectively, the DEAD-box domain of DDX41 and transmembrane region of STING. BTK phosphorylates DDX41, and its kinase activities are critical for STING-mediated IFN-β production. We show that Tyr364 and Tyr414 of DDX41 are critical for its recognition of AT-rich DNA and binding to STING, and tandem mass spectrometry identifies Tyr414 as the BTK phosphorylation site. Modeling studies further indicate that phospho-Tyr414 strengthens DDX41's interaction with STING. Hence, BTK plays a critical role in the activation of DDX41 helicase and STING signaling.

MeSH terms

Agammaglobulinaemia Tyrosine Kinase
Animals
Binding Sites
Cell Line
DEAD-box RNA Helicases / chemistry
DEAD-box RNA Helicases / genetics
DEAD-box RNA Helicases / metabolism*
DNA / chemistry
DNA / metabolism*
HEK293 Cells
Humans
Interferon Regulatory Factor-3 / metabolism
Interferon-beta / genetics
Interferon-beta / metabolism*
Membrane Proteins / chemistry
Membrane Proteins / genetics
Membrane Proteins / metabolism*
Mice
Mice, Inbred C57BL
Mice, Knockout
Molecular Dynamics Simulation
Parasitemia / mortality
Parasitemia / pathology
Parasitemia / veterinary
Phosphopeptides / analysis
Protein Binding
Protein Serine-Threonine Kinases / metabolism
Protein-Tyrosine Kinases / deficiency
Protein-Tyrosine Kinases / genetics
Protein-Tyrosine Kinases / metabolism*
Signal Transduction
Survival Rate

Substances

Interferon Regulatory Factor-3
Irf3 protein, mouse
Membrane Proteins
Phosphopeptides
Sting1 protein, mouse
Interferon-beta
DNA
Tbk1 protein, mouse
Protein-Tyrosine Kinases
Agammaglobulinaemia Tyrosine Kinase
BTK protein, human
Btk protein, mouse
Protein Serine-Threonine Kinases
DDX41 protein, mouse
DEAD-box RNA Helicases