Phenotypic Mutation 'Muskatenwein' (pdf version)
AlleleMuskatenwein
Mutation Type missense
Chromosome7
Coordinate141,307,176 bp (GRCm39)
Base Change T ⇒ A (forward strand)
Gene Muc2
Gene Name mucin 2
Synonym(s) 2010015E03Rik
Chromosomal Location 141,276,583-141,308,428 bp (+) (GRCm39)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] This gene encodes a member of the mucin protein family. Mucins are high molecular weight glycoproteins produced by many epithelial tissues. The protein encoded by this gene is secreted and forms an insoluble mucous barrier that protects the gut lumen. The protein polymerizes into a gel of which 80% is composed of oligosaccharide side chains by weight. The protein features a central domain containing tandem repeats rich in threonine and proline that varies between 50 and 115 copies in different individuals. Downregulation of this gene has been observed in patients with Crohn disease and ulcerative colitis. [provided by RefSeq, Oct 2016]
PHENOTYPE: Homozygotes for a point mutation have soft feces at weaning and develop diarrhea associated with malapsorption syndrome. Homozygous null mutants pass blood in their feces at 6 months, and 65% of null mutants have intestinal tumors at 1 year. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_023566, MGI: 1339364

MappedYes 
Amino Acid Change Isoleucine changed to Asparagine
Institutional SourceBeutler Lab
Gene Model not available
AlphaFold no structure available at present
SMART Domains Protein: ENSMUSP00000026590
Gene: ENSMUSG00000025515
AA Change: I336N

DomainStartEndE-ValueType
C8 1 63 1.65e-11 SMART
VWC 120 188 5.48e-2 SMART
VWC 229 293 2.38e-11 SMART
Blast:VWD 299 363 4e-17 BLAST
CT 380 463 3.6e-35 SMART
Predicted Effect unknown
Predicted Effect
Meta Mutation Damage Score Not available question?
Is this an essential gene? Probably nonessential (E-score: 0.099) question?
Phenotypic Category Autosomal Semidominant
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance 100% 
Alleles Listed at MGI

All alleles(7) : Targeted, knock-out(1) Targeted, other(2) Chemically induced(4)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
Eeyore APN 7 141693356 missense probably benign 0.35
kenny APN 7 nonsense
Winnie APN 7 141286029 missense probably damaging 1.00
IGL01303:Muc2 APN 7 141306132 missense probably benign
IGL01482:Muc2 APN 7 141307797 missense probably damaging 0.96
IGL01875:Muc2 APN 7 141306477 missense probably damaging 0.99
IGL02088:Muc2 APN 7 141305241 missense probably damaging 1.00
IGL02415:Muc2 APN 7 141305609 nonsense probably null
IGL02548:Muc2 APN 7 141305594 missense probably damaging 1.00
IGL02836:Muc2 APN 7 141300450 unclassified probably benign
IGL03196:Muc2 APN 7 141301367 missense probably damaging 0.97
nomoco UTSW 7 141307456 missense probably damaging 1.00
Schlendrian UTSW 7 141281925 missense probably damaging 1.00
Seco UTSW 7 141284976 missense probably damaging 1.00
BB001:Muc2 UTSW 7 141281631 missense probably damaging 1.00
BB011:Muc2 UTSW 7 141281631 missense probably damaging 1.00
E0370:Muc2 UTSW 7 141282598 missense probably damaging 1.00
R0127:Muc2 UTSW 7 141302691 missense probably benign 0.00
R0179:Muc2 UTSW 7 141302708 missense probably damaging 1.00
R0201:Muc2 UTSW 7 141699185 frame shift probably null
R0299:Muc2 UTSW 7 141306466 missense probably damaging 1.00
R0547:Muc2 UTSW 7 141699185 frame shift probably null
R0699:Muc2 UTSW 7 141306037 missense probably damaging 1.00
R0900:Muc2 UTSW 7 141699185 frame shift probably null
R1348:Muc2 UTSW 7 141699185 frame shift probably null
R1466:Muc2 UTSW 7 141302711 missense probably damaging 1.00
R1466:Muc2 UTSW 7 141302711 missense probably damaging 1.00
R1625:Muc2 UTSW 7 141283405 missense probably damaging 1.00
R2010:Muc2 UTSW 7 141287444 missense probably damaging 0.99
R2149:Muc2 UTSW 7 141699185 frame shift probably null
R2163:Muc2 UTSW 7 141699185 frame shift probably null
R3008:Muc2 UTSW 7 141281347 missense possibly damaging 0.93
R3110:Muc2 UTSW 7 141299225 unclassified probably benign
R3112:Muc2 UTSW 7 141299225 unclassified probably benign
R3424:Muc2 UTSW 7 141279595 missense probably damaging 0.99
R3786:Muc2 UTSW 7 141283590 missense probably benign 0.01
R3854:Muc2 UTSW 7 141308081 missense probably damaging 1.00
R3964:Muc2 UTSW 7 141286233 missense probably benign 0.17
R3965:Muc2 UTSW 7 141286233 missense probably benign 0.17
R3966:Muc2 UTSW 7 141286233 missense probably benign 0.17
R3973:Muc2 UTSW 7 141300541 unclassified probably benign
R3974:Muc2 UTSW 7 141300541 unclassified probably benign
R3976:Muc2 UTSW 7 141300541 unclassified probably benign
R4327:Muc2 UTSW 7 141281577 missense probably damaging 0.96
R4694:Muc2 UTSW 7 141306082 missense probably damaging 1.00
R4764:Muc2 UTSW 7 141299345 missense possibly damaging 0.88
R4769:Muc2 UTSW 7 141286260 critical splice donor site probably null
R4798:Muc2 UTSW 7 141307877 missense probably benign 0.01
R4900:Muc2 UTSW 7 141303280 missense probably benign 0.32
R5383:Muc2 UTSW 7 141307456 missense probably damaging 1.00
R5489:Muc2 UTSW 7 141305169 missense probably benign 0.00
R5615:Muc2 UTSW 7 141277446 missense probably damaging 1.00
R5856:Muc2 UTSW 7 141299381 unclassified probably benign
R5919:Muc2 UTSW 7 141281171 missense probably damaging 0.97
R5953:Muc2 UTSW 7 141287951 missense probably damaging 0.96
R5979:Muc2 UTSW 7 141305143 missense probably damaging 0.99
R5979:Muc2 UTSW 7 141283493 splice site probably null
R6175:Muc2 UTSW 7 141282875 missense probably damaging 1.00
R6213:Muc2 UTSW 7 141305151 missense probably damaging 1.00
R6281:Muc2 UTSW 7 141306140 missense probably damaging 1.00
R6321:Muc2 UTSW 7 141287397 missense probably benign 0.28
R6390:Muc2 UTSW 7 141305883 missense probably damaging 0.97
R6485:Muc2 UTSW 7 141300473 unclassified probably benign
R6582:Muc2 UTSW 7 141282941 missense probably benign 0.00
R6683:Muc2 UTSW 7 141305214 missense probably benign 0.38
R6896:Muc2 UTSW 7 141306432 missense possibly damaging 0.48
R6906:Muc2 UTSW 7 141284976 missense probably damaging 1.00
R6924:Muc2 UTSW 7 141284077 missense possibly damaging 0.87
R7040:Muc2 UTSW 7 141305194 missense unknown
R7222:Muc2 UTSW 7 141290758 missense
R7251:Muc2 UTSW 7 141278965 missense possibly damaging 0.91
R7282:Muc2 UTSW 7 141306481 missense
R7315:Muc2 UTSW 7 141276645 missense probably damaging 0.99
R7421:Muc2 UTSW 7 141301863 missense
R7556:Muc2 UTSW 7 141307439 missense
R7651:Muc2 UTSW 7 141290750 missense
R7710:Muc2 UTSW 7 141287452 missense possibly damaging 0.92
R7776:Muc2 UTSW 7 141290942 missense
R7813:Muc2 UTSW 7 141282543 splice site probably null
R7843:Muc2 UTSW 7 141281662 missense probably benign 0.03
R7869:Muc2 UTSW 7 141303471 missense
R7924:Muc2 UTSW 7 141281631 missense probably damaging 1.00
R7993:Muc2 UTSW 7 141308173 missense
R8053:Muc2 UTSW 7 141284575 missense probably benign 0.01
R8068:Muc2 UTSW 7 141298422 missense
R8099:Muc2 UTSW 7 141299175 splice site probably null
R8192:Muc2 UTSW 7 141305215 missense
R8194:Muc2 UTSW 7 141290801 missense
R8545:Muc2 UTSW 7 141306130 missense unknown
R8701:Muc2 UTSW 7 141281850 missense probably damaging 1.00
R8883:Muc2 UTSW 7 141287469 missense probably damaging 0.98
R8894:Muc2 UTSW 7 141280758 missense probably damaging 1.00
R8905:Muc2 UTSW 7 141279643 missense probably benign 0.00
R9024:Muc2 UTSW 7 141287936 missense probably damaging 0.98
R9032:Muc2 UTSW 7 141287058 missense probably damaging 1.00
R9085:Muc2 UTSW 7 141287058 missense probably damaging 1.00
R9091:Muc2 UTSW 7 141290816 missense
R9104:Muc2 UTSW 7 141286224 missense probably damaging 1.00
R9114:Muc2 UTSW 7 141287983 nonsense probably null
R9270:Muc2 UTSW 7 141290816 missense
R9297:Muc2 UTSW 7 141302759 missense
R9325:Muc2 UTSW 7 141298559 missense
R9354:Muc2 UTSW 7 141307157 missense
R9386:Muc2 UTSW 7 141279389 missense probably damaging 1.00
R9529:Muc2 UTSW 7 141287453 missense possibly damaging 0.55
R9550:Muc2 UTSW 7 141308242 missense probably damaging 1.00
R9583:Muc2 UTSW 7 141300559 missense
R9607:Muc2 UTSW 7 141305190 missense
R9646:Muc2 UTSW 7 141276643 missense probably benign
R9651:Muc2 UTSW 7 141288014 missense probably damaging 0.99
R9774:Muc2 UTSW 7 141285811 missense probably benign
R9784:Muc2 UTSW 7 141280785 nonsense probably null
Z1176:Muc2 UTSW 7 141300451 missense
Z1177:Muc2 UTSW 7 141298531 missense
Mode of Inheritance Autosomal Semidominant
Local Stock Live Mice, Sperm
MMRRC Submission 032797-UCD
Last Updated 2017-09-13 3:38 PM by Diantha La Vine
Record Created 2009-11-09 12:00 AM
Record Posted 2010-03-31
Other Mutations in This Stock Stock #: C9142 Run Code: SLD00193
Coding Region Coverage: 1x: 84.8% 3x: 59.3%
Validation Efficiency: 72/88

GeneSubstitutionChr/LocMutationPredicted EffectZygosity
Adnp A to G 2: 168,026,327 (GRCm39) S323P probably damaging Het
Cc2d2a A to G 5: 43,892,799 (GRCm39) probably benign Homo
Chrm5 A to G 2: 112,310,556 (GRCm39) F187L probably damaging Het
Fmnl3 T to C 15: 99,235,508 (GRCm39) probably null Het
Klf7 C to T 1: 64,118,316 (GRCm39) A94T possibly damaging Het
Nf1 C to T 11: 79,447,557 (GRCm39) R2433C probably damaging Het
Pdlim3 A to T 8: 46,349,869 (GRCm39) M60L probably benign Het
Vwde T to C 6: 13,168,053 (GRCm39) probably benign Homo
Phenotypic Description

The semidominant Muskatenwein phenotype was identified among ENU-mutagenized G3 mice in a screen for mutants with susceptibility to dextran sulfate sodium (DSS)-induced colitis (DSS-induced Colitis Screen).  When challenged with a low dose of DSS in their drinking water (1%), the index mouse (#48) showed severe bleeding and increased weight loss relative to wild type C57BL/6J mice on day 6 and day 7 of DSS exposure (Figure 1).    

Nature of Mutation
Whole genome sequencing of a heterozygous Muskatenwein mouse using the SOLiD technique identified a T to A transversion at base pair 148939344 on Chromosome 7 located in the Muc2 gene using NCBI m37 mouse assembly (Build 37.1). The mutation was confirmed using standard Sanger sequencing (Figure 2), and corresponds to nucleotide 6506 of the Muc2 transcript in exon 44 of 47 coding exons (1)
 
6490 ATCAACAACCAGCTCATCTCCTCGGTCTCCAAC
2167 -I--N--N--Q--L--I--S--S--V--S--N-
 
The mutated nucleotide is indicated in red lettering, and causes an isoleucine to asparagine change at amino acid 2172 according to the NP_076055 record and amino acid 2533 according to Uniprot record Q80Z19
Illustration of Mutations in
Gene & Protein
Protein Prediction
Figure 3.  Domain structure of Mucin2. The Muskatenwein mutation alters an isoleucine residue located near the C-terminal end of the protein between the C domain and the cysteine knot (CK) domain. S=Signal sequence; D=D domains (homology to VWF mediates trimerization); CR=Cystein-rich domain; TR= Tandem repeat domain (heavily O glycosylated); GDPH=GDPH autocatalytic proteolytic site; B=B domain (homology to VWF); C= C domain (homology to VWF); CK= Cysteine-knot domain (homology to VWF mediates dimerization). This image is interactive. Click on the image to view other mutations found in Mucin2 (red). Click on the mutations for more specific information.

The Muskatenwein mutation alters an isoleucine residue located near the C-terminal end of the protein between the C domain and the cysteine knot (CK) domain (Figure 3). 

 

For more information about Muc2, please see the record for Schlendrian.

Putative Mechanism
The DSS-induced colitis found in Muskatenwein animals remains unmapped, but this phenotype is similar if less severe to the phenotypes seen in mice carrying missense Muc2 mutations (see the records for Schlendrian, Winnie and Eeyore) (2) and suggests that the Muc2 mutation found in Muskatenwein mice is indeed the phenotype causing mutation. Like other missense mutations in Muc2 and unlike Muc2-/- animals (2,3), the Muskatenwein phenotype is semidominant suggesting the presence of aberrant protein in these animals. Aberrant MUC2 proteins display abnormal oligomerization and accumulation in the endoplasmic reticulum (ER) leading to ER stress, triggering of the unfolded protein response (UPR), subsequent inflammation and goblet cell apoptosis (2).
 
Although the isoleucine residue altered by the Muskatenwein mutation does not occur in any defined domain, the phenotype of these mice suggests that it is important for MUC2 function. Furthermore, this residue is highly conserved and occurs in a conserved region of the MUC2 protein. The Muskatenwein missense mutation is predicted to be possibly damaging by the PolyPhen(see report). The D3 and CK domains of MUC2 have been implicated in trimerization and dimerization of MUC2 molecules through the formation of disulfide bonds (4-6), and it is possible that altering amino acids located near the CK domain may also affect the biosynthesis and assembly of MUC2 proteins. 
Primers Primers cannot be located by automatic search.
Genotyping
Muskatenwein genotyping is performed by amplifying the region containing the mutation using PCR, followed by sequencing of the amplified region to detect the single nucleotide change.
 
Primers for PCR amplification
Musk(F): 5’- AGCAGACCCATTGCATCATCGAG -3’
Musk(R): 5’- TCCTTCATGACGGAGACAGCAGAG -3’
 
PCR program
1) 94°C             2:00
2) 94°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               ∞
 
Primers for sequencing
Musk_seq(F): 5’- GATACTCATTCCTGGAGCTAGGAC -3’
Musk_seq(R): 5’- TTTGAGGAACTGAGTCCCAAC -3’
 
The following sequence of 1390 nucleotides (NCBI Mouse Genome Build 37.1, Chromosome 7 bases 148938628 to 148940017) is amplified:
 
agcagaccca ttgcatcatc gaggggccca agcagcagta cattattctg aaggtgtgtg
cgcttctggc tccacccaca agctaggcag ccactgaggg caaggctagg atgacttcgg
gcttgagtca tcctaaatcc cacatagcat atttgggtgg ctaaaagcaa catatatggt
tgctccactt ctggagctct gaagtgtaag atcaaggctg gcagatgctg tgaggagtcc
ttcctgcctc ttctttgggc tccagccaat tttggtttgt gtttgaatca cctctgatgt
agtagtgtcc ttctatctga gcctgtgcct cctcttctgt ctcttataag gatactcatt
cctggagcta ggacctcctt catgtagggt gacctcatat ccagattatt ttcttgagct
tttcaaagat catttcccca ctcagagtca ctttctagtt cacaagtcag acctaggtta
gagatttctg tgttatgtaa ccacagtagc tatctccaag actgtaacgg atgccacttg
cccagtgact ctggacctcc tttctgtggc gctttgcctc ccaggtcttc ctgggctggc
ctctggcgta cacctgctca tcctcattct gttgcgtcca cagcctgggg agattcacaa
aaaccccagc aacaagtgca ccttcttcag ctgcatgaaa atcaacaacc agctcatctc
ctcggtctcc aacatcacct gtcccgactt caacccaagt gattgtgttt cagtgagtgg
gaccctatgg ccctgtccat gtatgcgttg ccaggagcag ggaaccttaa tgcaccagct
acctggtttc cagggggctt gttccacacc ctcctgtagg acagccctta tgcatcccat
gacaagaagg ggcactcact tgctgggggt gctctggact cctttcttac tctcctttct
cttcagggct ccatcacata catgcctaat ggctgctgta agacatgtga gtacggctgg
ggaaatgttg ggtgatgggt gagttttact cagagccttg aggggaagat gcaggacagg
atgcttcagg actgtgtctc ctacactcat gcagatgtat aggaatctga gtctccttcc
agggagaggc tgggtggagc ccagctgcat gttgggactc agttcctcaa aggaaagtgg
gactcagtta agggctagag acatgctcgg agaaggaaga agaaatggca ccagagaagg
agagggaggg gaagtgggga gagagaggga aggggcaagg ggagctagct agctagagtc
atttttcccc cacaggcatc cctcaaaacc agaccagagt cccttgctct gctgtctccg
tcatgaagga
 
PCR primer binding sites are underlined; sequencing primer binding sites are highlighted in gray; the mutated T is shown in red text.
References
Science Writers Nora G. Smart
Illustrators Nora G. Smart, Diantha La Vine
AuthorsKatharina Brandl, Bruce Beutler
Edit History
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