Ubiquitin Ligase TRIM62 Regulates CARD9-Mediated Anti-fungal Immunity and Intestinal Inflammation

Zhifang Cao; Kara L Conway; Robert J Heath; Jason S Rush; Elizaveta S Leshchiner; Zaida G Ramirez-Ortiz; Natalia B Nedelsky; Hailiang Huang; Aylwin Ng; Agnès Gardet; Shih-Chin Cheng; Alykhan F Shamji; John D Rioux; Cisca Wijmenga; Mihai G Netea; Terry K Means; Mark J Daly; Ramnik J Xavier

doi:10.1016/j.immuni.2015.10.005

Ubiquitin Ligase TRIM62 Regulates CARD9-Mediated Anti-fungal Immunity and Intestinal Inflammation

Immunity. 2015 Oct 20;43(4):715-26. doi: 10.1016/j.immuni.2015.10.005.

Authors

Zhifang Cao¹, Kara L Conway², Robert J Heath², Jason S Rush³, Elizaveta S Leshchiner³, Zaida G Ramirez-Ortiz⁴, Natalia B Nedelsky¹, Hailiang Huang⁵, Aylwin Ng², Agnès Gardet¹, Shih-Chin Cheng⁶, Alykhan F Shamji³, John D Rioux⁷, Cisca Wijmenga⁸, Mihai G Netea⁶, Terry K Means⁴, Mark J Daly⁵, Ramnik J Xavier⁹

Affiliations

¹ Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
² Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
³ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA.
⁴ Center for Immunology and Inflammatory Diseases and Division of Rheumatology, Allergy, and Immunology, Massachusetts General Hospital and Harvard Medical School, Charlestown, MA 02129, USA.
⁵ Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA; Analytic and Translational Genetics Unit, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA.
⁶ Department of Internal Medicine and Radboud Center for Infectious Diseases, Radboud University Nijmegen Medical Center, Nijmegen 6525 GA, the Netherlands.
⁷ Research Center, Montreal Heart Institute and Université de Montréal, QC H1T 1C8, Canada.
⁸ Department of Genetics, University Medical Center Groningen, University of Groningen, Groningen T9700 RB, the Netherlands.
⁹ Gastrointestinal Unit and Center for the Study of Inflammatory Bowel Disease, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Center for Computational and Integrative Biology, Massachusetts General Hospital and Harvard Medical School, Boston, MA 02114, USA; Broad Institute of MIT and Harvard, Cambridge, MA 02142, USA. Electronic address: xavier@molbio.mgh.harvard.edu.

Abstract

CARD9 is a central component of anti-fungal innate immune signaling via C-type lectin receptors, and several immune-related disorders are associated with CARD9 alterations. Here, we used a rare CARD9 variant that confers protection against inflammatory bowel disease as an entry point to investigating CARD9 regulation. We showed that the protective variant of CARD9, which is C-terminally truncated, acted in a dominant-negative manner for CARD9-mediated cytokine production, indicating an important role for the C terminus in CARD9 signaling. We identified TRIM62 as a CARD9 binding partner and showed that TRIM62 facilitated K27-linked poly-ubiquitination of CARD9. We identified K125 as the ubiquitinated residue on CARD9 and demonstrated that this ubiquitination was essential for CARD9 activity. Furthermore, we showed that similar to Card9-deficient mice, Trim62-deficient mice had increased susceptibility to fungal infection. In this study, we utilized a rare protective allele to uncover a TRIM62-mediated mechanism for regulation of CARD9 activation.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Adjuvants, Immunologic / pharmacology
Animals
CARD Signaling Adaptor Proteins / chemistry
CARD Signaling Adaptor Proteins / deficiency
CARD Signaling Adaptor Proteins / genetics
CARD Signaling Adaptor Proteins / physiology*
Candidiasis, Invasive / genetics
Candidiasis, Invasive / immunology*
Colitis / chemically induced
Colitis / genetics
Colitis / prevention & control
Cytokines / biosynthesis
Dendritic Cells / immunology
Dendritic Cells / metabolism
Genes, Dominant
Genetic Predisposition to Disease
HEK293 Cells
HeLa Cells
Humans
Inflammatory Bowel Diseases / genetics
Mice
Mice, 129 Strain
Mice, Knockout
Protein Interaction Mapping
Protein Isoforms / chemistry
Protein Isoforms / genetics
Protein Isoforms / physiology
Protein Processing, Post-Translational
Protein Structure, Tertiary
Receptors, Angiotensin / chemistry
Receptors, Angiotensin / deficiency
Receptors, Angiotensin / physiology*
Receptors, Endothelin / chemistry
Receptors, Endothelin / deficiency
Receptors, Endothelin / physiology*
Recombinant Fusion Proteins / metabolism
Signal Transduction
Specific Pathogen-Free Organisms
Tripartite Motif Proteins
Ubiquitin-Protein Ligases / chemistry
Ubiquitin-Protein Ligases / physiology*
Ubiquitination

Substances

Adjuvants, Immunologic
CARD Signaling Adaptor Proteins
CARD9 protein, human
Card9 protein, mouse
Cytokines
Dear1 protein, mouse
Protein Isoforms
Receptors, Angiotensin
Receptors, Endothelin
Recombinant Fusion Proteins
Tripartite Motif Proteins
TRIM62 protein, human
Ubiquitin-Protein Ligases

Abstract

Publication types

MeSH terms

Substances

Grants and funding