|Coordinate||59,549,354 bp (GRCm38)|
|Base Change||A ⇒ G (forward strand)|
|Gene Name||NLR family, pyrin domain containing 3|
|Synonym(s)||Cias1, cryopyrin, Pypaf1, NALP3, Mmig1|
|Chromosomal Location||59,541,568-59,566,955 bp (+)|
|MGI Phenotype||Strain: 3686871
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.
|Amino Acid Change||Arginine changed to Glycine|
|Institutional Source||Beutler Lab|
R586G in Ensembl: ENSMUSP00000078440 (fasta)
|Gene Model||not available|
|Predicted Effect||possibly damaging
PolyPhen 2 Score 0.586 (Sensitivity: 0.88; Specificity: 0.91)
|Phenotypic Category||immune system, NALP3 inflammasome signaling defect|
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Semidominant|
|Local Stock||Sperm, gDNA|
|Last Updated||03/02/2017 12:13 PM by Katherine Timer|
|Record Created||06/05/2010 8:25 PM by Hua Huang|
The ND6 phenotype was initially identified among G3 mice (G5801 and G5806) homozygous for mutations induced by N-ethyl-N-nitrosourea (ENU) and tested in the NALP3 Inflammasome Screen. Peritoneal macrophages isolated from these mice secreted reduced amounts of the proinflammatory cytokine interleukin (IL)-1β in response to priming with lipopolysaccharide (LPS) followed by nigericin treatment (Figure 1). Macrophages from G5801 produced normal levels of tumor necrosis factor (TNF)-α in response to LPS stimulation alone, suggesting that signaling from the Toll-like receptor 4 (TLR4), which senses LPS, was unimpaired.
|Nature of Mutation|
The Nlrp3 gene was directly sequenced as a candidate gene and an A to G transition was found at position 1982 of the Nlrp3 transcript in exon 4 of 10 total exons using Genbank record NM_145827. The mutation is located in the third coding exon.
The mutated nucleotide is indicated in red lettering, and causes an arginine to glycine substitution at residue 586 of the NLRP3 protein.
Please see the record for ND1 for more information about Nlrp3.
NLRP3 is able to oligomerize through its NBD domain and assemble into large caspase-1-activating multiprotein complexes termed inflammasomes upon the detection of pathogenic or other danger signals in the cytoplasm. A large variety of agents have been shown to activate the NLRP3 inflammasome, and NLRP3 plays an important role in the innate immune response.
A total of 93 disease-associated mutations have been found in humans in the NLRP3 gene according to the Infevers database, an online database for autoinflammatory mutations (1-3). Rather than causing immunodeficiencies, these are activating mutations that cause autoinflammatory disorders including Muckle-Wells syndrome (MWS; #191900), familial cold autoinflammatory syndrome (FCAS1; OMIM #120100), and chronic infantile neurologic cutaneous and articular syndrome (CINCA; #607115), also known as NOMID for neonatal onset multisystem inflammatory disease (4-6). Collectively, these diseases are known as cryopyrin associated periodic syndrome (CAPS). Interestingly, the vast majority of these activating missense mutations are located in exon 3, which encodes the NBD and surrounding sequences. These mutations are thought to either allow the protein the ability to permanently bind to and hydrolyze ATP or unable to bind to the inhibitory LRRs that prevent self-oligomerization (7-8). However, the ND6 mutation, which is located in the same exon, appears to be an inactivating mutation and renders macrophages unresponsive to NLRP3-activating stimuli. The mechanism by which the ND6 mutation affects NLRP3 function is unknown.
|Primers||Primers cannot be located by automatic search.|
ND6 genotyping is performed by amplifying the region containing the mutation using PCR, followed by sequencing of the amplified region to detect the single nucleotide transition.
ND6(F): 5’- AGGCTGATCCAAGAGAATGAGGTCC -3’
ND6(R): 5’- AACACACTGAACCTGGTCCAAGGG -3’
1) 95°C 2:00
2) 95°C 0:30
3) 56°C 0:30
4) 72°C 1:00
5) repeat steps (2-4) 29X
6) 72°C 7:00
7) 4°C 8
Primers for sequencing
ND6 _seq(F): 5'- ATCTTCTGAACCGAGACGTG -3'
ND6 _seq(R): 5’- AGCGTTTTGACCCTATGACAG -3’
The following sequence of 939 nucleotides (NCBI Mouse Genome Build 37.1, Chromosome 11, bases 59,362,277 to 59,363,215) is amplified:
aggctgatcc aagagaatga ggtcctcttt accatgtgct tcatccccct ggtctgctgg
attgtgtgca cggggctaaa gcaacagatg gagaccggga agagcctggc ccagacctcc
aagaccacta cggccgtcta cgtcttcttc ctttccagcc tgctgcaatc ccgggggggc
attgaggagc atctcttctc tgactaccta caggggctct gttcactggc tgcggatgga
atttggaacc agaaaatcct atttgaggag tgtgatctgc ggaagcacgg cctgcagaag
actgacgtct ccgctttcct gaggatgaac gtgttccaga aggaagtgga ctgcgagaga
ttctacagct tcagccacat gactttccag gagttcttcg ctgctatgta ctatttgctg
gaagaggagg cagaggggga gaccgtgagg aaaggaccag gaggttgttc agatcttctg
aaccgagacg tgaaggtcct actagaaaat tacggcaagt ttgaaaaagg ctatctgatt
tttgttgtcc gattcctctt tggccttgta aaccaggaga gaacctctta tttggagaag
aaactaagtt gcaagatctc tcagcaagtc agactggaac tactgaagtg gattgaagtg
aaagccaagg ccaagaagct gcagtggcag cccagccaac tggaactgtt ctactgcctg
tacgagatgc aggaggaaga ctttgtgcag agtgccatgg accactttcc caaaattgag
atcaacctct ctaccagaat ggaccacgtg gtttcctcct tttgtattaa gaactgtcat
agggtcaaaa cgctttccct gggttttttt cacaactcgc ccaaggagga agaagaagag
aggagaggag gtcgaccctt ggaccaggtt cagtgtgtt
Primer binding sites are underlined; sequencing primer binding sites are highlighted in gray; the mutated A is indicated in red.
1. Milhavet, F., Cuisset, L., Hoffman, H. M., Slim, R., El-Shanti, H., Aksentijevich, I., Lesage, S., Waterham, H., Wise, C., Sarrauste de Menthiere, C., and Touitou, I. (2008) The Infevers Autoinflammatory Mutation Online Registry: Update with New Genes and Functions. Hum. Mutat. 29, 803-808.
2. Touitou, I., Lesage, S., McDermott, M., Cuisset, L., Hoffman, H., Dode, C., Shoham, N., Aganna, E., Hugot, J. P., Wise, C., Waterham, H., Pugnere, D., Demaille, J., and Sarrauste de Menthiere, C. (2004) Infevers: An Evolving Mutation Database for Auto-Inflammatory Syndromes. Hum. Mutat. 24, 194-198.
3. Sarrauste de Menthiere, C., Terriere, S., Pugnere, D., Ruiz, M., Demaille, J., and Touitou, I. (2003) INFEVERS: The Registry for FMF and Hereditary Inflammatory Disorders Mutations. Nucleic Acids Res. 31, 282-285.
4. Dode, C., Le Du, N., Cuisset, L., Letourneur, F., Berthelot, J. M., Vaudour, G., Meyrier, A., Watts, R. A., Scott, D. G., Nicholls, A., Granel, B., Frances, C., Garcier, F., Edery, P., Boulinguez, S., Domergues, J. P., Delpech, M., and Grateau, G. (2002) New Mutations of CIAS1 that are Responsible for Muckle-Wells Syndrome and Familial Cold Urticaria: A Novel Mutation Underlies both Syndromes. Am. J. Hum. Genet. 70, 1498-1506.
5. Feldmann, J., Prieur, A. M., Quartier, P., Berquin, P., Certain, S., Cortis, E., Teillac-Hamel, D., Fischer, A., and de Saint Basile, G. (2002) Chronic Infantile Neurological Cutaneous and Articular Syndrome is Caused by Mutations in CIAS1, a Gene Highly Expressed in Polymorphonuclear Cells and Chondrocytes. Am. J. Hum. Genet. 71, 198-203.
6. Hoffman, H. M., Rosengren, S., Boyle, D. L., Cho, J. Y., Nayar, J., Mueller, J. L., Anderson, J. P., Wanderer, A. A., and Firestein, G. S. (2004) Prevention of Cold-Associated Acute Inflammation in Familial Cold Autoinflammatory Syndrome by Interleukin-1 Receptor Antagonist. Lancet. 364, 1779-1785.
7. Neven, B., Callebaut, I., Prieur, A. M., Feldmann, J., Bodemer, C., Lepore, L., Derfalvi, B., Benjaponpitak, S., Vesely, R., Sauvain, M. J., Oertle, S., Allen, R., Morgan, G., Borkhardt, A., Hill, C., Gardner-Medwin, J., Fischer, A., and de Saint Basile, G. (2004) Molecular Basis of the Spectral Expression of CIAS1 Mutations Associated with Phagocytic Cell-Mediated Autoinflammatory Disorders CINCA/NOMID, MWS, and FCU. Blood. 103, 2809-2815.
|Science Writers||Nora G. Smart|
|Authors||Hua Huang, Bruce Beutler|