Phenotypic Mutation 'unmodulated' (pdf version)
Alleleunmodulated
Mutation Type intron
Chromosome5
Coordinate140,897,997 bp (GRCm38)
Base Change A ⇒ T (forward strand)
Gene Card11
Gene Name caspase recruitment domain family, member 11
Synonym(s) 2410011D02Rik, BIMP3, CARMA1, 0610008L17Rik
Chromosomal Location 140,858,745-140,986,337 bp (-) (GRCm39)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] The protein encoded by this gene belongs to the membrane-associated guanylate kinase (MAGUK) family, a class of proteins that functions as molecular scaffolds for the assembly of multiprotein complexes at specialized regions of the plasma membrane. This protein is also a member of the CARD protein family, which is defined by carrying a characteristic caspase-associated recruitment domain (CARD). This protein has a domain structure similar to that of CARD14 protein. The CARD domains of both proteins have been shown to specifically interact with BCL10, a protein known to function as a positive regulator of cell apoptosis and NF-kappaB activation. When expressed in cells, this protein activated NF-kappaB and induced the phosphorylation of BCL10. [provided by RefSeq, Jul 2008]
PHENOTYPE: Mice homozygous for a targeted null mutation exhibit defects in antigen receptor signalling in both T and B lymphocytes. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_175362; MGI: 1916978

MappedYes 
Amino Acid Change
Institutional SourceAustralian Phenomics Network
Gene Model not available
AlphaFold no structure available at present
SMART Domains Protein: ENSMUSP00000082941
Gene: ENSMUSG00000036526

DomainStartEndE-ValueType
Pfam:CARD 23 109 1.3e-23 PFAM
coiled coil region 176 440 N/A INTRINSIC
low complexity region 475 487 N/A INTRINSIC
low complexity region 535 549 N/A INTRINSIC
low complexity region 615 625 N/A INTRINSIC
PDZ 674 755 2.73e-1 SMART
Blast:SH3 776 838 1e-10 BLAST
low complexity region 839 850 N/A INTRINSIC
low complexity region 920 934 N/A INTRINSIC
SCOP:d1kjwa2 970 1149 1e-18 SMART
Blast:GuKc 973 1139 1e-102 BLAST
Predicted Effect probably benign
Meta Mutation Damage Score Not available question?
Is this an essential gene? Non Essential (E-score: 0.000) question?
Phenotypic Category Autosomal Recessive
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance 100% 
Alleles Listed at MGI

All alleles(6) : Targeted, knock-out(3) Targeted, other(1) Chemically induced(2)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00961:Card11 APN 5 140885464 missense probably damaging 0.97
IGL01645:Card11 APN 5 140863778 missense probably benign 0.00
IGL01731:Card11 APN 5 140868057 missense possibly damaging 0.89
IGL01782:Card11 APN 5 140913481 start codon destroyed probably null 0.02
IGL01935:Card11 APN 5 140869301 missense possibly damaging 0.62
IGL01991:Card11 APN 5 140899133 missense possibly damaging 0.63
IGL02447:Card11 APN 5 140892679 missense possibly damaging 0.93
IGL02583:Card11 APN 5 140863881 missense probably benign 0.10
IGL03255:Card11 APN 5 140884086 missense possibly damaging 0.73
Ace UTSW 5 140888632 missense possibly damaging 0.70
Caravaggio UTSW 5 140899064 missense probably damaging 1.00
Dealer UTSW 5 140871632 missense probably damaging 1.00
Dogs UTSW 5 140867755 critical splice donor site probably null
Face UTSW 5 140886732 missense probably damaging 1.00
hubei UTSW 5 140892522 missense probably damaging 0.96
king UTSW 5 140876835 splice site probably benign
may UTSW 5 140862250 nonsense probably null
Poker UTSW 5 140863837 missense probably benign
Sharp UTSW 5 140862180 missense possibly damaging 0.93
Tumnus UTSW 5 140871700 missense possibly damaging 0.75
unmodulated2 UTSW 5 140869537 splice site probably null
PIT4243001:Card11 UTSW 5 140894359 missense possibly damaging 0.95
PIT4486001:Card11 UTSW 5 140862163 missense probably damaging 1.00
PIT4531001:Card11 UTSW 5 140892415 missense probably damaging 0.99
R0046:Card11 UTSW 5 140894279 missense possibly damaging 0.92
R0285:Card11 UTSW 5 140872856 missense probably damaging 1.00
R0452:Card11 UTSW 5 140866125 missense probably benign 0.01
R1486:Card11 UTSW 5 140862274 missense probably benign
R1710:Card11 UTSW 5 140888660 nonsense probably null
R1733:Card11 UTSW 5 140892388 missense possibly damaging 0.88
R1817:Card11 UTSW 5 140871315 missense probably benign 0.00
R1818:Card11 UTSW 5 140871315 missense probably benign 0.00
R2027:Card11 UTSW 5 140892522 missense probably damaging 0.96
R2436:Card11 UTSW 5 140868117 missense possibly damaging 0.89
R2904:Card11 UTSW 5 140874888 missense probably benign 0.09
R3706:Card11 UTSW 5 140872890 missense probably damaging 0.99
R3708:Card11 UTSW 5 140872890 missense probably damaging 0.99
R4778:Card11 UTSW 5 140869537 splice site probably null
R4877:Card11 UTSW 5 140871632 missense probably damaging 1.00
R4889:Card11 UTSW 5 140871700 missense possibly damaging 0.75
R4910:Card11 UTSW 5 140860169 missense probably damaging 1.00
R5011:Card11 UTSW 5 140862275 missense possibly damaging 0.93
R5257:Card11 UTSW 5 140862180 missense possibly damaging 0.93
R5258:Card11 UTSW 5 140862180 missense possibly damaging 0.93
R5682:Card11 UTSW 5 140888666 nonsense probably null
R5754:Card11 UTSW 5 140885524 missense probably damaging 0.99
R5873:Card11 UTSW 5 140894393 missense probably damaging 1.00
R6184:Card11 UTSW 5 140884033 missense probably damaging 1.00
R6792:Card11 UTSW 5 140899064 missense probably damaging 1.00
R6825:Card11 UTSW 5 140863837 missense probably benign
R7008:Card11 UTSW 5 140859148 missense probably damaging 1.00
R7291:Card11 UTSW 5 140886825 missense probably damaging 1.00
R7376:Card11 UTSW 5 140883993 missense probably benign 0.01
R7526:Card11 UTSW 5 140899184 splice site probably null
R7683:Card11 UTSW 5 140881781 missense probably benign
R7730:Card11 UTSW 5 140871751 missense probably damaging 0.96
R7813:Card11 UTSW 5 140885419 missense probably damaging 1.00
R7831:Card11 UTSW 5 140859167 missense possibly damaging 0.61
R7911:Card11 UTSW 5 140867755 critical splice donor site probably null
R8154:Card11 UTSW 5 140886732 missense probably damaging 1.00
R8224:Card11 UTSW 5 140888632 missense possibly damaging 0.70
R8272:Card11 UTSW 5 140875794 missense probably damaging 1.00
R8714:Card11 UTSW 5 140899147 missense possibly damaging 0.67
R8715:Card11 UTSW 5 140871315 missense probably benign 0.00
R9065:Card11 UTSW 5 140894297 missense probably damaging 1.00
R9211:Card11 UTSW 5 140869375 missense probably benign 0.16
R9215:Card11 UTSW 5 140866154 missense possibly damaging 0.64
R9269:Card11 UTSW 5 140892516 missense probably damaging 0.99
R9385:Card11 UTSW 5 140871276 missense probably benign 0.44
R9421:Card11 UTSW 5 140869462 missense probably damaging 0.97
R9424:Card11 UTSW 5 140894395 missense probably damaging 1.00
R9444:Card11 UTSW 5 140894393 missense probably damaging 1.00
V7732:Card11 UTSW 5 140862250 nonsense probably null
X0067:Card11 UTSW 5 140871347 missense possibly damaging 0.60
Z1177:Card11 UTSW 5 140883996 missense probably benign 0.43
Mode of Inheritance Autosomal Recessive
Local Stock None
Repository

Australian PhenomeBank: 2

Last Updated 2019-04-16 7:12 PM by Diantha La Vine
Record Created 2011-01-10 5:07 PM by Eva Marie Y. Moresco
Record Posted 2011-01-10
Phenotypic Description
Figure 1. Flow cytometric analysis of splenic B cells in homozygous unmodulated (un/un) and wild type mice. Numbers indicate the percentage of gated population. Dashed lines indicate the peak intensity of CD21 in wild type controls. Figure reproduced from (1).

The unmodulated phenotype was first identified among G3 mice in a flow cytometric screen for blood lymphocyte abnormalities (1).  Several mice in one pedigree exhibited an unusually high level of surface IgM on circulating IgD+ B cells (Figure 1).  Further characterization also demonstrated higher levels of surface CD21 on these cells (Figure 1), and CD23 on follicular B cells.  Unmodulated homozygotes displayed normal frequencies of pre-B and immature B cells in the bone marrow, normal frequencies of T and B cells in lymph node and spleen, normal maturation of thymic T cell subsets, and normal numbers of circulating CD4+ and CD8+ T cells in the lymph nodes and spleen.  The number of IgD+ mature follicular B cells in the spleen was also normal.  In the peritoneal cavity, the B1 cell subset (IgM+ CD5+ CD23Mac1+) was nearly absent although the number of B2 cells was unaffected.

Figure 2. Defective T-dependent antibody responses in unmodulated mice. (A) Basal serum antibody levels in wild type and homozygous unmodulated (un/un) mice. (B) DNP-specific antibody levels 9 and 8 days after soluble DNP-Ficoll and alum-precipitated DNP-CGG immunization, respectively. (C) Bordatella perussis-specific and CGG-specific IgG2a and IgG1 antibody responses 16 days after combined alum-precipitated CGG plus B. pertussis immunization. Figure reproduced from (1).

Sera from unmodulated mice had 50% less IgM and 90% less IgG3 than wild type control mice.  Serum IgG2B and IgG1 levels were not significantly different from wild type (Figure 2A).  Compared to wild type control mice, unmodulated animals showed little increase in serum IgM and no increase in serum IgG3 titers to the DNP hapten following immunization with DNP-Ficoll, a T cell-independent type 2 antigen (Figure 2B). There was also a dramatic decrease in the primary antibody response to immunization with the T cell-dependent antigen, chicken γ globulin (CGG; Figure 2B), and poor formation of germinal centers.  Th2 type antibody responses to alum-precipitated DNP-CGG were reduced, and Th1 type antibody responses to heat-killed Bordetella pertussis were undetectable 16 days after combined immunization (Figure 2C).

Figure 3. Selective defects in B cell responses. (A) Division of CFSE-labeled wild type and homosygous unmodulated B cells stimulated with anti-IgM or LPS for 72 hours. Shaded histogram shows undivided cells in parallel cultures with IL-4 alone. (B) BCR-induced degradation of IκBα measured by Western blotting. Results are representative of three experiments. (C) Phosphorylation of ERK and JNK measured in purified B cells by Western blotting with anti-phospho-ERK1/2 or anti-phospho-JNK antibodies. Blots were reprobed for β-actin to confirm equal loading. Shown below is densitometric intensity of phospho-JNK relative to unstimulated cells and corrected for β-actin signal. Results are representative of three experiments. Figure reproduced from (1).

In response to crosslinking B cell antigen receptors with IgM antibodies, unmodulated B cells displayed greatly reduced proliferation compared to wild type B cells, most undergoing only one division by 72 hours (Figure 3A).  Proliferation induced by antibody to the CD40 receptor was also reduced.  NF-κB (Figure 3B) and JNK activation (Figure 3C) were selectively impaired in unmodulated B cells, whereas intracellular calcium elevation, and NFAT and ERK (Figure 3C) activation occurred normally.  The proliferative response to TLR4 activation with lipopolysaccharide (LPS) was normal in unmodulated B cells (Figure 3A).

Figure 4. Selective defects in T cell responses. (A) DNA synthesis after 48 hr in unfractionated splenocytes from wild type (filled symbols) and mutant (unfilled symbols) mice stimulated with the indicated concentrations of plate-bound anti-TCR in the presence (triangles) or absence (circles) of anti-CD28. (B) Division of CFSE-labeled wild type and mutant CD4+ T cells after 72 hr in unfractionated cultures stimulated with 10 µm/ml anti-TCR cultured separately (left) or with mutant and wild type mized 1:1 and cocultured (right). Dashed lines indicate modal CFSE in undivided cells. Figure reproduced from (1).

In contrast to the severe proliferative defect in B cells, unmodulated T cells enlarged and engaged in DNA synthesis normally during the first 48 hours of culture when stimulated by crosslinking their T cell antigen receptors (Figure 4A).  However, they were refractory to costimulation by the addition of CD28 antibodies (Figure 4A), and failed to divide more than one or two times (Figure 4B).  NF-κB activation was also diminished in response to CD28 costimulatory signals.

Figure 5. Spontaneous atopy in unmodulated mice. (A) Serun IgE concentration in mice of the indicated genotypes and age groups. (B) Representative histology of wild-type and mutant mouse skin with atopic dermatitis, stained with hematoxylin and eosin (H&E) or Alcian blue to reveal mast cells (arrowhead). Figure reproduced from (1).

The majority of homozygous mutant mice but none of their heterozygous or wild type littermates began to develop high serum levels of IgE, teary eyes, and erythematous ears at 10–20 weeks of age (Figure 5A).  Their ears became intensely itchy, progressing to dermatitis of the ear and neck.  Histologically, the inflamed regions of skin exhibited hyperkeratosis, extensive mast cell infiltrates, mononuclear cells, and occasional eosinophils (Figure 5B).

The cellular defects of unmodulated mice were found to be cell-intrinsic.

Nature of Mutation

The unmodulated mutation was mapped to Chromosome 5, and corresponds to a T to A transversion at position 1235 of the Card11 transcript, in exon 7 of 25 total exons.

1219 GCCATCTTGGACATCCTGGAACATGACCGGAAG

300  -A--I--L--D--I--L--E--H--D--R--K-

Figure 6. Domain structure of CARMA1. The unmodulated mutation results in a leucine to glutamine substitution at position 305 within the coiled-coil region. Four predicted α-helices are represented in pink. This image is interactive. Other mutations found in CARMA1 are noted. Click on each mututation for more specific information.

The mutated nucleotide is indicated in red lettering, and results in a leucine to glutamine substitution at position 305 of the encoded CARMA1 protein (Figure 6).

Illustration of Mutations in
Gene & Protein
Putative Mechanism

The phenotype of homozygous unmodulated mice is similar to those of other Card11 mutants including Card11-/- (2;3), Card11king/king(4), and a knock-in mutant expressing a CARD-deleted CARMA1 protein (5).  The unmodulated mutation lies within the coiled-coil domain that mediates the localization and constitutive oligomerization of CARMA1 (6).  The mutated residue is a conserved leucine in the heptad repeats of the predicted α-helical coil.  Western blotting with an antiserum to Carma-1 revealed normal levels of the protein in mutant B cells. The change to polar glutamine from nonpolar leucine would be predicted to disrupt the coiled-coil structure and interfere with binding partner interactions.

Please see the record for king for information about Card11.

Primers Primers cannot be located by automatic search.
Genotyping

Genotyping protocol available from the Australian PhenomeBank: Carma-1 (Unmodulated).

References
Science Writers Eva Marie Y. Moresco
Illustrators Diantha La Vine
AuthorsJesse E. Jun, Lauren E. Wilson, Carola G. Vinuesa, Sylvie Lesage, Mathieu Blery, Lisa A. Miosge, Matthew C. Cook, Edyta M. Kucharska, Hiromitsu Hara, Josef M. Penninger, Heather Domashenz, Nancy A. Hong, Richard J. Glynne, Keats A. Nelms, Christopher C. Goodnow
Edit History
2011-01-17 12:26 PM (current)
2011-01-17 12:25 PM
2011-01-11 9:35 AM
2011-01-11 9:33 AM
2011-01-10 5:29 PM
2011-01-10 5:24 PM
2011-01-10 5:21 PM
2011-01-10 5:21 PM
2011-01-10 5:21 PM