Phenotypic Mutation 'Lps4' (pdf version)
AlleleLps4
Mutation Type missense
Chromosome4
Coordinate66,759,379 bp (GRCm39)
Base Change G ⇒ A (forward strand)
Gene Tlr4
Gene Name toll-like receptor 4
Synonym(s) Lps, lipopolysaccharide response, Rasl2-8
Chromosomal Location 66,745,788-66,765,338 bp (+) (GRCm39)
MGI Phenotype FUNCTION: This gene belongs to the evolutionarily-conserved Toll-like receptor family, whose members are type-1 transmembrane proteins that are involved in innate immunity. Toll-like receptors are characterized by an extracellular leucine-rich repeat domain that functions in ligand recognition and an intracellular toll/interleukin-1 receptor-like domain that is crucial for signal transduction. The receptor encoded by this gene mediates the innate immune response to bacterial lipopolysaccharide, a major component of the outer membrane of Gram-negative bacteria, through synthesis of pro-inflammatory cytokines and chemokines. In addition, this protein can recognize other pathogens from Gram-negative and Gram-positive bacteria as well as viral components. Mice deficient in this gene display a number of immune response-related phenotypes including hyporesponsiveness to bacterial lipopolysaccharide and increased levels of respiratory syncytial virus compared to controls. [provided by RefSeq, Sep 2015]
PHENOTYPE: Homozygotes for spontaneous or targeted mutations are hyporesponsive to bacterial lipopolysaccharide and more susceptible to infection by gram negative bacteria. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_021297; MGI: 96824

MappedYes 
Amino Acid Change Glycine changed to Aspartic acid
Institutional SourceBeutler Lab
Gene Model not available
AlphaFold Q9QUK6
PDB Structure Crystal structure of mouse TLR4 and mouse MD-2 complex [X-RAY DIFFRACTION]
Crystal structure of mouse TLR4/MD-2/lipid IVa complex [X-RAY DIFFRACTION]
Crystal structure of mouse TLR4/MD-2/LPS complex [X-RAY DIFFRACTION]
SMART Domains Protein: ENSMUSP00000045770
Gene: ENSMUSG00000039005
AA Change: G724D

DomainStartEndE-ValueType
LRR 76 99 7.36e0 SMART
LRR 100 123 1.86e0 SMART
LRR 173 196 8.24e0 SMART
LRR 370 401 4.33e1 SMART
LRR 468 492 2.54e2 SMART
LRR 493 516 1.86e2 SMART
LRR 517 540 1.67e2 SMART
LRR 541 563 1.92e2 SMART
LRRCT 576 626 4.74e-3 SMART
transmembrane domain 636 658 N/A INTRINSIC
TIR 671 816 7.3e-39 SMART
low complexity region 822 833 N/A INTRINSIC
Predicted Effect probably damaging

PolyPhen 2 Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
(Using ENSMUST00000048096)
SMART Domains Protein: ENSMUSP00000102988
Gene: ENSMUSG00000039005

DomainStartEndE-ValueType
PDB:3VQ2|B 22 86 2e-38 PDB
SCOP:d1m0za_ 27 86 4e-6 SMART
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 Semidominant
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance 100% 
Alleles Listed at MGI
All alleles(8) : Targeted, knock-out(1) Spontaneous(5) Chemically induced(2)
Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL01120:Tlr4 APN 4 66758662 missense probably benign 0.01
IGL01343:Tlr4 APN 4 66752124 splice site probably benign
IGL01669:Tlr4 APN 4 66759504 missense possibly damaging 0.48
IGL01875:Tlr4 APN 4 66757726 missense probably damaging 1.00
IGL02138:Tlr4 APN 4 66759202 missense probably damaging 0.99
IGL02244:Tlr4 APN 4 66752298 critical splice donor site probably null
IGL02793:Tlr4 APN 4 66757681 missense probably damaging 1.00
IGL03269:Tlr4 APN 4 66759033 missense probably damaging 1.00
IGL03288:Tlr4 APN 4 66757990 missense probably damaging 0.99
bugsy UTSW 4 66757491 nonsense probably null
Cruyff UTSW 4 66758563 missense probably damaging 1.00
don_knotts UTSW 4 66759409 missense probably damaging 1.00
Guardiola UTSW 4 66757540 missense probably damaging 1.00
Lops UTSW 4 66752117 splice site probably null
lps3 UTSW 4 66759334 missense probably damaging 1.00
milquetoast UTSW 4 66757681 missense probably damaging 1.00
salvador UTSW 4 66758443 missense probably damaging 0.99
R0449:Tlr4 UTSW 4 66757857 missense probably damaging 0.99
R0481:Tlr4 UTSW 4 66746153 missense probably benign 0.05
R0576:Tlr4 UTSW 4 66757732 missense probably benign 0.00
R0827:Tlr4 UTSW 4 66752117 splice site probably null
R1488:Tlr4 UTSW 4 66757786 missense probably damaging 1.00
R1490:Tlr4 UTSW 4 66757611 missense possibly damaging 0.56
R1522:Tlr4 UTSW 4 66757933 missense possibly damaging 0.80
R1616:Tlr4 UTSW 4 66757717 missense probably damaging 1.00
R1681:Tlr4 UTSW 4 66759342 missense probably damaging 1.00
R1738:Tlr4 UTSW 4 66759313 missense probably benign 0.19
R1888:Tlr4 UTSW 4 66759409 missense probably damaging 1.00
R1888:Tlr4 UTSW 4 66759409 missense probably damaging 1.00
R1929:Tlr4 UTSW 4 66757681 missense probably damaging 1.00
R1982:Tlr4 UTSW 4 66759272 missense probably benign 0.40
R1998:Tlr4 UTSW 4 66758707 missense probably damaging 1.00
R2186:Tlr4 UTSW 4 66758220 missense possibly damaging 0.63
R2305:Tlr4 UTSW 4 66758338 missense probably damaging 1.00
R3011:Tlr4 UTSW 4 66757491 nonsense probably null
R3420:Tlr4 UTSW 4 66757773 missense probably benign 0.37
R3422:Tlr4 UTSW 4 66757773 missense probably benign 0.37
R3818:Tlr4 UTSW 4 66759553 missense probably benign 0.00
R4212:Tlr4 UTSW 4 66758563 missense probably damaging 1.00
R4213:Tlr4 UTSW 4 66758563 missense probably damaging 1.00
R4417:Tlr4 UTSW 4 66757540 missense probably damaging 1.00
R4630:Tlr4 UTSW 4 66757477 missense probably benign 0.44
R4735:Tlr4 UTSW 4 66759435 missense probably damaging 1.00
R5191:Tlr4 UTSW 4 66759616 missense probably damaging 0.96
R5613:Tlr4 UTSW 4 66759122 missense possibly damaging 0.94
R5705:Tlr4 UTSW 4 66752217 missense probably damaging 1.00
R5726:Tlr4 UTSW 4 66758652 missense probably benign
R6021:Tlr4 UTSW 4 66759103 missense probably damaging 1.00
R6159:Tlr4 UTSW 4 66758070 missense possibly damaging 0.92
R6227:Tlr4 UTSW 4 66758832 missense probably benign
R7139:Tlr4 UTSW 4 66758520 missense probably benign 0.06
R7199:Tlr4 UTSW 4 66759430 missense probably damaging 0.99
R7220:Tlr4 UTSW 4 66758188 missense probably benign
R7337:Tlr4 UTSW 4 66758191 missense possibly damaging 0.86
R7487:Tlr4 UTSW 4 66842659 missense probably benign 0.00
R7638:Tlr4 UTSW 4 66758443 missense probably damaging 0.99
R7773:Tlr4 UTSW 4 66757836 missense probably damaging 1.00
R7814:Tlr4 UTSW 4 66759316 missense probably damaging 1.00
R7897:Tlr4 UTSW 4 66758058 missense probably benign 0.07
R8044:Tlr4 UTSW 4 66746084 missense probably benign 0.01
R8062:Tlr4 UTSW 4 66758087 missense probably benign 0.00
R8080:Tlr4 UTSW 4 66757713 missense probably damaging 1.00
R8446:Tlr4 UTSW 4 66757673 missense probably damaging 0.98
R8916:Tlr4 UTSW 4 66847268 missense probably benign 0.06
R9100:Tlr4 UTSW 4 66758518 missense probably benign 0.08
R9415:Tlr4 UTSW 4 66746160 critical splice donor site probably null
R9562:Tlr4 UTSW 4 66759522 missense possibly damaging 0.80
R9565:Tlr4 UTSW 4 66759522 missense possibly damaging 0.80
R9752:Tlr4 UTSW 4 66757912 missense probably benign 0.02
X0064:Tlr4 UTSW 4 66758377 missense probably damaging 0.99
Z1088:Tlr4 UTSW 4 66847319 missense probably benign 0.01
Mode of Inheritance Autosomal Semidominant
Local Stock Sperm, gDNA
MMRRC Submission 031073-UCD
Last Updated 2016-12-09 10:31 AM by Katherine Timer
Record Created unknown
Record Posted 2009-04-16
Phenotypic Description

The Lps4 phenotype was identified in an ENU-mutagenized G3 mouse, which had a reduced interleukin (IL)-1β response to LPS/nigericin (Figure 1A).  Subsequent testing of isolated peritoneal macrophages from additional G3 animals, including siblings of the original index mouse, found a defect in tumor necrosis factor (TNF)-α production in response to the toll-like receptor 4 (TLR4) ligand lipopolysaccharide (LPS).  TNF-α production is normal in response to all other TLR ligands tested (TLR Signaling Screen; Figure 1B).  The original index mouse has been genotyped and is heterozygous for the mutation.  

Nature of Mutation
The Tlr4 gene on Chromosome 4 of Lps4 mice was sequenced, and a G to A transition was identified in exon 3 (of 3 total exons) at position 2192 of the Tlr4 transcript.
 
2176 AACATCATCCAGGAAGGCTTCCACAAGAGCCGG
719  -N--I--I--Q--E--G--F--H--K--S--R-
 
The mutated nucleotide is indicated in red lettering, and results in a glycine to aspartic acid change at amino acid 724 of the TLR4 protein.
Illustration of Mutations in
Gene & Protein
Protein Prediction
 
Figure 2. Protein and domain structure of TLR4. A) Schematic representation of TLR9 based on crystalized structures of mouse TLR3 LRR (PBD 3CIG) and human TLR2 TIR (1FYW) domains. The residue affected by the Lps4 mutation is highlighted. 3D image was created using UCSF Chimera. B) TLR4 is an 835 amino acid protein with an extracellur domain (pink) of leucine rich repeats (LRR), a short transmembrane domain and a cytoplasmic Toll/Interleukin-1 receptor (TIR) domain. The Lps4 mutation (red asterisk) results in a glycine to aspartic acid change at amino acid 724 of the TLR4 protein. This image is interactive. Click on the image to view other mutations found in TLR4 (red). Click on the mutations for more specific information.
The Lps4 mutation results in a glycine to aspartic acid change at amino acid 724, which is located towards the end of the αβ helix of the TLR4 TIR domain (Figures 2 and 3).  The TIR domain participates in protein-protein interactions, but a specific role for the αβ helix has not been described.  This residue is conserved amongst TLR4 homologues, but is not conserved with other TLR proteins.  It is likely the substitution of a charged amino acid for a glycine at this position disrupts helical formation as the previous amino acid is also charged. 
 
Please see the record for lps3 for information about Tlr4.
Primers Primers cannot be located by automatic search.
Genotyping
Lps4 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
Lps4(F): 5’- TCTTGGCTTGACCAGTCTCAACAC -3’
Lps4(R): 5’- TTGTCCTCCCATTCCAGGTAGGTG -3’
 
PCR program
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              ∞
 
Primers for sequencing
Lps4_seq(F1): 5’- GGGGTATTTGACACCCTCCATAG -3’
Lps4_seq(F2): 5’- ACAATCATCAGTGTGTCAGTGG -3’
Lps4_seq(R): 5’- GTTTCTGCTAAGAAGGCGATAC -3’
 
The following sequence of 999 nucleotides (from Genbank genomic region NC_000070 for linear DNA sequence of Tlr4) is amplified:
 
12556                 tcttg gcttgaccag tctcaacaca ttaaaaatgg ctggcaattc
12601 tttcaaagac aacacccttt caaatgtctt tgcaaacaca acaaacttga cattcctgga
12661 tctttctaaa tgtcaattgg aacaaatatc ttggggggta tttgacaccc tccatagact
12721 tcaattatta aatatgagtc acaacaatct attgtttttg gattcatccc attataacca
12781 gctgtattcc ctcagcactc ttgattgcag tttcaatcgc atagagacat ctaaaggaat
12841 actgcaacat tttccaaaga gtctagcctt cttcaatctt actaacaatt ctgttgcttg
12901 tatatgtgaa catcagaaat tcctgcagtg ggtcaaggaa cagaagcagt tcttggtgaa
12961 tgttgaacaa atgacatgtg caacacctgt agagatgaat acctccttag tgttggattt
13021 taataattct acctgttata tgtacaagac aatcatcagt gtgtcagtgg tcagtgtgat
13081 tgtggtatcc actgtagcat ttctgatata ccacttctat tttcacctga tacttattgc
13141 tggctgtaaa aagtacagca gaggagaaag catctatgat gcatttgtga tctactcgag
13201 tcagaatgag gactgggtga gaaatgagct ggtaaagaat ttagaagaag gagtgccccg
13261 ctttcacctc tgccttcact acagagactt tattcctggt gtagccattg ctgccaacat
13321 catccaggaa ggcttccaca agagccggaa ggttattgtg gtagtgtcta gacactttat
13381 tcagagccgt tggtgtatct ttgaatatga gattgctcaa acatggcagt ttctgagcag
13441 ccgctctggc atcatcttca ttgtccttga gaaggttgag aagtccctgc tgaggcagca
13501 ggtggaattg tatcgccttc ttagcagaaa cacctacctg gaatgggagg acaa
 
PCR primer binding sites are underlined; sequencing primer binding sites are highlighted in gray; the mutated G is shown in red text.
Science Writers Nora G. Smart
Illustrators Diantha La Vine
AuthorsOwen Siggs, Bruce Beutler
Edit History
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