Phenotypic Mutation 'Park2' (pdf version)
AllelePark2
Mutation Type nonsense
Chromosome11
Coordinate59,455,954 bp (GRCm39)
Base Change C ⇒ T (forward strand)
Gene Nlrp3
Gene Name NLR family, pyrin domain containing 3
Synonym(s) Mmig1, Cias1, NALP3, cryopyrin, Pypaf1
Chromosomal Location 59,432,395-59,457,781 bp (+) (GRCm39)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] This gene encodes a pyrin-like protein containing a pyrin domain, a nucleotide-binding site (NBS) domain, and a leucine-rich repeat (LRR) motif. This protein interacts with the apoptosis-associated speck-like protein PYCARD/ASC, which contains a caspase recruitment domain, and is a member of the NALP3 inflammasome complex. This complex functions as an upstream activator of NF-kappaB signaling, and it plays a role in the regulation of inflammation, the immune response, and apoptosis. Mutations in this gene are associated with familial cold autoinflammatory syndrome (FCAS), Muckle-Wells syndrome (MWS), chronic infantile neurological cutaneous and articular (CINCA) syndrome, and neonatal-onset multisystem inflammatory disease (NOMID). Multiple alternatively spliced transcript variants encoding distinct isoforms have been identified for this gene. Alternative 5' UTR structures are suggested by available data; however, insufficient evidence is available to determine if all of the represented 5' UTR splice patterns are biologically valid. [provided by RefSeq, Oct 2008]
PHENOTYPE: Mice homozygous for null mutations exhibit attenuated inflammatory responses related to decrease secretion of IL-1beta and IL-18. Mice heterozygous for activating mutations suffer from autoinflammatory attacks that lead to organ failure and death before weaning. [provided by MGI curators]
Accession Number

Ncbi RefSeq: NM_145827.3; MGI: 2653833

MappedYes 
Amino Acid Change Arginine changed to Stop codon
Institutional SourceBeutler Lab
Gene Model predicted gene model for protein(s): [ENSMUSP00000078440] [ENSMUSP00000098707]
AlphaFold Q8R4B8
SMART Domains Protein: ENSMUSP00000078440
Gene: ENSMUSG00000032691
AA Change: R917*

DomainStartEndE-ValueType
PYRIN 4 87 6.39e-33 SMART
FISNA 135 206 1.45e-22 SMART
Pfam:NACHT 216 385 6.7e-52 PFAM
low complexity region 533 539 N/A INTRINSIC
low complexity region 688 697 N/A INTRINSIC
LRR_RI 737 764 1.07e-9 SMART
LRR 766 793 5.13e1 SMART
LRR 794 821 3.86e-7 SMART
LRR 823 850 1.62e0 SMART
LRR 851 878 3.39e-3 SMART
LRR 880 907 1.2e2 SMART
LRR 908 935 2.24e-3 SMART
LRR 937 964 2.16e2 SMART
LRR 965 992 8.73e-6 SMART
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000098707
Gene: ENSMUSG00000032691
AA Change: R917*

DomainStartEndE-ValueType
PYRIN 4 87 6.39e-33 SMART
FISNA 135 206 1.45e-22 SMART
Pfam:NACHT 216 385 6.7e-52 PFAM
low complexity region 533 539 N/A INTRINSIC
low complexity region 688 697 N/A INTRINSIC
LRR_RI 737 764 1.07e-9 SMART
LRR 766 793 5.13e1 SMART
LRR 794 821 3.86e-7 SMART
LRR 823 850 1.62e0 SMART
LRR 851 878 3.39e-3 SMART
LRR 880 907 1.2e2 SMART
LRR 908 935 2.24e-3 SMART
LRR 937 964 2.16e2 SMART
LRR 965 992 8.73e-6 SMART
Predicted Effect probably null
Meta Mutation Damage Score 0.9754 question?
Is this an essential gene? Probably nonessential (E-score: 0.076) question?
Phenotypic Category Autosomal Semidominant
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance  
Alleles Listed at MGI

All alleles(13) : Targeted(9) Chemically induced(4)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00421:Nlrp3 APN 11 59456769 missense probably damaging 0.99
IGL00573:Nlrp3 APN 11 59455942 missense possibly damaging 0.93
IGL01025:Nlrp3 APN 11 59442713 missense probably benign 0.21
IGL01637:Nlrp3 APN 11 59440204 missense probably damaging 0.99
IGL02010:Nlrp3 APN 11 59440361 missense probably benign
IGL02334:Nlrp3 APN 11 59455909 missense probably benign
IGL02417:Nlrp3 APN 11 59456849 unclassified probably benign
IGL02578:Nlrp3 APN 11 59439227 missense probably damaging 1.00
IGL02710:Nlrp3 APN 11 59456802 missense probably damaging 0.99
IGL02816:Nlrp3 APN 11 59446608 missense probably benign 0.03
IGL03157:Nlrp3 APN 11 59440372 missense possibly damaging 0.80
IGL03334:Nlrp3 APN 11 59439842 missense probably damaging 1.00
Flogiston UTSW 11 59449274 missense probably benign 0.00
nd1 UTSW 11 59456800 missense probably benign 0.45
Nd14 UTSW 11 59446701 missense possibly damaging 0.89
Nd3 UTSW 11 59456800 missense probably benign 0.45
nd5 UTSW 11 59456705 missense probably benign 0.01
nd6 UTSW 11 59440180 missense probably damaging 1.00
nd7 UTSW 11 59446701 missense possibly damaging 0.89
Nd9 UTSW 11 59440180 missense probably damaging 1.00
Park3 UTSW 11 59456676 missense probably benign 0.02
Park4 UTSW 11 59440357 missense probably benign 0.19
Park5 UTSW 11 59439302 missense probably damaging 0.99
Park6 UTSW 11 59439862 missense probably damaging 1.00
Park7 UTSW 11 59438836 nonsense probably null
Park8 UTSW 11 59457025 missense probably benign 0.19
R0008:Nlrp3 UTSW 11 59449274 missense probably benign 0.00
R0008:Nlrp3 UTSW 11 59449274 missense probably benign 0.00
R0052:Nlrp3 UTSW 11 59455954 nonsense probably null
R0362:Nlrp3 UTSW 11 59439623 missense possibly damaging 0.49
R0416:Nlrp3 UTSW 11 59446750 splice site probably benign
R0649:Nlrp3 UTSW 11 59439368 missense possibly damaging 0.83
R0740:Nlrp3 UTSW 11 59439082 missense probably benign 0.01
R0863:Nlrp3 UTSW 11 59456676 missense probably benign 0.02
R1300:Nlrp3 UTSW 11 59446594 missense possibly damaging 0.86
R1414:Nlrp3 UTSW 11 59440357 missense probably benign 0.19
R1622:Nlrp3 UTSW 11 59439302 missense probably damaging 0.99
R1654:Nlrp3 UTSW 11 59433949 missense probably benign 0.03
R1715:Nlrp3 UTSW 11 59434177 missense probably damaging 1.00
R1754:Nlrp3 UTSW 11 59449228 missense possibly damaging 0.80
R1837:Nlrp3 UTSW 11 59439742 missense probably benign 0.00
R1905:Nlrp3 UTSW 11 59439862 missense probably damaging 1.00
R2281:Nlrp3 UTSW 11 59439962 missense possibly damaging 0.70
R4296:Nlrp3 UTSW 11 59440487 missense possibly damaging 0.89
R4305:Nlrp3 UTSW 11 59438836 nonsense probably null
R4540:Nlrp3 UTSW 11 59442725 missense possibly damaging 0.83
R4591:Nlrp3 UTSW 11 59440048 missense probably benign 0.00
R4816:Nlrp3 UTSW 11 59439127 missense probably benign 0.32
R4913:Nlrp3 UTSW 11 59440064 missense probably benign 0.09
R4970:Nlrp3 UTSW 11 59439554 missense probably damaging 1.00
R5051:Nlrp3 UTSW 11 59457025 missense probably benign 0.19
R5112:Nlrp3 UTSW 11 59439554 missense probably damaging 1.00
R5185:Nlrp3 UTSW 11 59455910 missense probably benign 0.05
R5417:Nlrp3 UTSW 11 59439889 missense probably damaging 1.00
R5709:Nlrp3 UTSW 11 59446574 nonsense probably null
R5869:Nlrp3 UTSW 11 59438960 missense probably damaging 1.00
R5898:Nlrp3 UTSW 11 59437678 missense probably benign 0.00
R5953:Nlrp3 UTSW 11 59437617 missense probably benign
R5979:Nlrp3 UTSW 11 59439797 missense probably benign 0.06
R6359:Nlrp3 UTSW 11 59439392 missense probably damaging 0.97
R6723:Nlrp3 UTSW 11 59456018 missense probably damaging 1.00
R7261:Nlrp3 UTSW 11 59439272 missense possibly damaging 0.83
R7349:Nlrp3 UTSW 11 59438912 missense probably damaging 1.00
R7388:Nlrp3 UTSW 11 59455892 missense probably benign 0.00
R7715:Nlrp3 UTSW 11 59433829 splice site probably null
R7916:Nlrp3 UTSW 11 59442689 missense probably benign 0.00
R8222:Nlrp3 UTSW 11 59439614 missense probably damaging 0.98
R8360:Nlrp3 UTSW 11 59440229 missense probably benign 0.02
R8390:Nlrp3 UTSW 11 59442616 missense possibly damaging 0.47
R8550:Nlrp3 UTSW 11 59440097 missense probably damaging 1.00
R8738:Nlrp3 UTSW 11 59440216 missense probably benign 0.00
R8940:Nlrp3 UTSW 11 59455870 missense probably benign 0.26
R8990:Nlrp3 UTSW 11 59439584 missense probably damaging 0.99
R9324:Nlrp3 UTSW 11 59434141 missense probably damaging 1.00
R9673:Nlrp3 UTSW 11 59440148 missense probably damaging 1.00
RF031:Nlrp3 UTSW 11 59449378 frame shift probably null
RF040:Nlrp3 UTSW 11 59449378 frame shift probably null
Z1088:Nlrp3 UTSW 11 59442686 missense possibly damaging 0.67
Mode of Inheritance Autosomal Semidominant
Local Stock
MMRRC Submission 036813-MU
Last Updated 2019-01-29 1:24 PM by Diantha La Vine
Record Created 2012-12-17 10:50 PM by Hexin Shi
Record Posted 2013-03-04
Phenotypic Description
Figure 1. The Park2 mice exhibit attenuated inflammatory responses related to decrease secretion of IL-1β. IL-1β were determined by ELISA.  T1560, T1720, and T1559 (red dots) are Park2 mice.
Figure 2. Both homozygous and heterozygous Park2 mice exhibit attenuated IL-1β secretion in response to nigericin treatment.

The Park2 phenotype was initially identified among G3 mice for mutations induced by N-ethyl-N-nitrosourea (ENU) and tested in the NALP3 Inflammasome Screen. An IL-1β-specific ELISA determined that perintoneal macrophages isolated from Park2 mice have attenuated inflammatory responses related to decrease secretion of the proinflammatory cytokine interleukin (IL)-1β in response to priming with lipopolysaccharide (LPS) followed by nigericin treatment (Figure 1); heterozgous Park2 animals also show a defect in IL-1β secretion following nigericin treatment (Figure 2). Park2 animals produced normal levels of tumor necrosis factor (TNF)-α in response to LPS stimulation alone (not shown), suggesting that signaling from the Toll-like receptor 4 (TLR4), which senses LPS, was unimpaired.

Nature of Mutation

Whole genome sequencing using the SOLiD technique identified 31 mutations in the G1 mouse.  One of which was a mutation in Nlrp3. Mutations in Nlrp3 have been documented to cause defects in the NALP3 Inflammasome Screen (click on Figure 3 in "Protein Prediction" for our ENU-induced Nlrp3 mutations).  Capillary sequencing of Nlrp3 (n = 10 G3 mice) identified a C to T transition at base pair 59378630 (v37) on chromosome 11 in the GenBank genomic region NC_000077 encoding Nlrp3. The mutation corresponds to residue 2749 in the Nlrp3 cDNA (ENSMUST00000079476) in exon 9 of 11.

2737 CACCTCTATCTACGAAGCAATGCCCTT

913  -H--L--Y--L--R--S--N--A--L-

The mutated nucleotide is indicated in red lettering and results in substitution of arginine (R) 917 for a premature stop codon.

Illustration of Mutations in
Gene & Protein
Protein Prediction

Figure 2. Domain structure of NLRP3. Shown are the pyrin domain (PYD), NACHT domain, NACHT associated domain (NAD), and leucine-rich repeat (LRR) domain. Together, the NACHT and NAD domains are known as the nucleotide-binding domain (NBD). The Park2 mutation results in substitution of arginine 917 for a premature stop codon. This image is interactive. Other mutations found in NLRP3 are noted in red. Click on each mutation for more specific information.

The Park2 mutation encodes a premature stop codon within the LRR domain of NLRP3 (Figure 3). The LRR domain assists in protein-protein interactions and in NLRP3 this domain binds to the nucleotide-binding domain (NBD) to prevent oligomerization and protein activation (1). The NBD consists of the NACHT and NAD domains together.

For more information about Nlrp3, please see the record for ND1.

Putative Mechanism

The Park2 mutation results in coding of a premature stop codon at residue 917. It is possible that this mutation results in the loss of protein oligomerization and activation.  The Park2 mutation is in proximity to the ND1 and ND5 mutations in Nlrp3, both of which are proposed to be inactivating mutations. The phenotype of the Park2 mice suggests that the mutated protein, if expressed, is inactive or has reduced activity.  In addition, the loss of IL-1β secretion in the heterozygous mouse indicates that this mutation alters the function of the wild-type protein.

Primers PCR Primer
Park2_pcr_F: AAGTTGGAAGGATGCAGCTCGC
Park2_pcr_R: AATGAATGTTTGCAGTTGGCAGGAC

Genotyping

Park2 genotyping is performed by amplifying the region containing the mutation using PCR followed by sequencing of the amplified region to detect the nucleotide change.  The following primers were used for PCR amplification and sequencing:

PCR Primers

Park2_For: 5’- AAGTTGGAAGGATGCAGCTCGC -3’

Park2_Rev: 5’ -AATGAATGTTTGCAGTTGGCAGGAC -3’

Sequencing Primer

Park2_Seq_For: 5’ - GCAGTGACACACAATGTCTG -3’

PCR program

1) 94° C        2:00

2) 94° C        0:30

3) 57° C        0:30

4) 72° C        1:00

5) repeat steps (2-4) 29x

6) 72° C        7:00

7) 4° C            -

The following sequence of 449 nucleotides (from Genbank genomic region NC_000077.6 of the linear genomic sequence Mus musculus strain C57BL/6J chromosome 11, GRCm38.p1 C57BL/6J encoding Nlrp3) is amplified:

  1 aagttggaag gatgcagctc gcagtgacac acaatgtctg agagagcctg ggcactgagg

 61 actccttctc ttcttcctat aggttggtga attccggcct tacttcaatc tgttgttcag

121 ctctgacctc tgtgctcaaa accaaccaga acttcacaca cctctatcta cgaagcaatg

181 cccttggaga cacaggactc aggctcctct gtgaggggct tctgcacccg gactgtaaac

241 tacagatgct ggagtgagtt catgggctgc ctgaggtgat gggaacatgg ggtgtgaggc

301 aggccctgga gcaaccctga tgcaaatctg tgtctgagga gtgaggcgtt cttggtgtcc

361 attagtgaga aatgggaagt ttctagatgg tatacagctt aaaggctaag cccctgccac

421 cacagtcctg ccaactgcaa acattcatt

PCR primer binding sites are underlined; the sequencing primer binding site is highlighted; the mutated C is highlighted in red.

References
Science Writers Anne Murray
Illustrators Katherine Timer
AuthorsHexin Shi, Ying Wang, Bruce Beutler