|Coordinate||59,565,128 bp (GRCm38)|
|Base Change||C ⇒ T (forward strand)|
|Gene Name||NLR family, pyrin domain containing 3|
|Synonym(s)||Cias1, cryopyrin, Pypaf1, NALP3, Mmig1|
|Chromosomal Location||59,541,568-59,566,956 bp (+)|
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]
|Amino Acid Change||Arginine changed to Stop codon|
|Institutional Source||Beutler Lab|
|Gene Model||predicted gene model for protein(s): [ENSMUSP00000078440] [ENSMUSP00000098707]|
AA Change: R917*
|Predicted Effect||probably null|
AA Change: R917*
|Predicted Effect||probably null|
|Meta Mutation Damage Score||0.9754|
|Is this an essential gene?||Probably nonessential (E-score: 0.081)|
|Phenotypic Category||Autosomal Semidominant|
|Candidate Explorer Status||loading ...|
Linkage Analysis Data
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Semidominant|
|Last Updated||2019-01-29 1:24 PM by Diantha La Vine|
|Record Created||2012-12-17 10:50 PM by Hexin Shi|
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.
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
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.
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.
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:
Park2_For: 5’- AAGTTGGAAGGATGCAGCTCGC -3’
Park2_Rev: 5’ -AATGAATGTTTGCAGTTGGCAGGAC -3’
Park2_Seq_For: 5’ - GCAGTGACACACAATGTCTG -3’
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.
|Science Writers||Anne Murray|
|Authors||Hexin Shi, Ying Wang, Bruce Beutler|