Phenotypic Mutation 'gordo' (pdf version)
Allelegordo
Mutation Type missense
Chromosome4
Coordinate101,622,502 bp (GRCm39)
Base Change T ⇒ A (forward strand)
Gene Lepr
Gene Name leptin receptor
Synonym(s) leptin receptor gene-related protein, obl, Obr, Leprb, obese-like, Modb1, LEPROT, OB-RGRP
Chromosomal Location 101,574,601-101,672,549 bp (+) (GRCm39)
MGI Phenotype FUNCTION: [Summary is not available for the mouse gene. This summary is for the human ortholog.] The protein encoded by this gene belongs to the gp130 family of cytokine receptors that are known to stimulate gene transcription via activation of cytosolic STAT proteins. This protein is a receptor for leptin (an adipocyte-specific hormone that regulates body weight), and is involved in the regulation of fat metabolism, as well as in a novel hematopoietic pathway that is required for normal lymphopoiesis. Mutations in this gene have been associated with obesity and pituitary dysfunction. Alternatively spliced transcript variants encoding different isoforms have been described for this gene. It is noteworthy that this gene and LEPROT gene (GeneID:54741) share the same promoter and the first 2 exons, however, encode distinct proteins (PMID:9207021).[provided by RefSeq, Nov 2010]
PHENOTYPE: Homozygous mutants are hyperphagic, low-activity, poorly cold-adapted, sterile and have enhanced fat conversion. They are obese, hyperinsulinemic and, on certain strains, severely hyperglycemic. Heterozygotes are normal but resistant to prolonged fasting. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_146146, NM_010704, NM_001122899; MGI:104993

MappedYes 
Amino Acid Change Aspartic acid changed to Glutamic Acid
Institutional SourceBeutler Lab
Gene Model predicted gene model for protein(s): [ENSMUSP00000037385] [ENSMUSP00000099838] [ENSMUSP00000102534]
AlphaFold P48356
SMART Domains Protein: ENSMUSP00000037385
Gene: ENSMUSG00000057722
AA Change: D427E

DomainStartEndE-ValueType
transmembrane domain 7 29 N/A INTRINSIC
FN3 236 315 1.5e-5 SMART
Pfam:Lep_receptor_Ig 328 418 6.3e-23 PFAM
FN3 535 618 4.93e-1 SMART
FN3 641 721 3.25e1 SMART
FN3 736 818 2.35e0 SMART
transmembrane domain 838 860 N/A INTRINSIC
low complexity region 908 921 N/A INTRINSIC
low complexity region 1050 1065 N/A INTRINSIC
Predicted Effect probably damaging

PolyPhen 2 Score 0.972 (Sensitivity: 0.77; Specificity: 0.96)
(Using ENSMUST00000037552)
SMART Domains Protein: ENSMUSP00000099838
Gene: ENSMUSG00000057722
AA Change: D427E

DomainStartEndE-ValueType
transmembrane domain 7 29 N/A INTRINSIC
FN3 236 315 1.5e-5 SMART
Pfam:Lep_receptor_Ig 329 420 2.6e-29 PFAM
FN3 535 618 4.93e-1 SMART
FN3 641 721 3.25e1 SMART
FN3 736 818 2.35e0 SMART
transmembrane domain 838 860 N/A INTRINSIC
Predicted Effect probably damaging

PolyPhen 2 Score 0.972 (Sensitivity: 0.77; Specificity: 0.96)
(Using ENSMUST00000102777)
SMART Domains Protein: ENSMUSP00000102534
Gene: ENSMUSG00000057722
AA Change: D427E

DomainStartEndE-ValueType
transmembrane domain 7 29 N/A INTRINSIC
FN3 236 315 1.5e-5 SMART
Pfam:Lep_receptor_Ig 329 420 2.6e-29 PFAM
FN3 535 618 4.93e-1 SMART
FN3 641 721 3.25e1 SMART
FN3 736 818 2.35e0 SMART
transmembrane domain 838 860 N/A INTRINSIC
Predicted Effect probably damaging

PolyPhen 2 Score 0.972 (Sensitivity: 0.77; Specificity: 0.96)
(Using ENSMUST00000106921)
Meta Mutation Damage Score 0.2622 question?
Is this an essential gene? Non Essential (E-score: 0.000) question?
Phenotypic Category Unknown
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance  
Alleles Listed at MGI

All Mutations and Alleles(55) : Chemically induced (ENU)(11) Chemically induced (other)(2) Radiation induced(1) Spontaneous(15) Targeted(23) Transgenic(3)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00089:Lepr APN 4 101672232 missense probably benign
IGL01111:Lepr APN 4 101671852 missense possibly damaging 0.77
IGL01324:Lepr APN 4 101625265 missense probably benign 0.23
IGL01372:Lepr APN 4 101592774 missense possibly damaging 0.67
IGL01626:Lepr APN 4 101590731 missense probably benign 0.10
IGL01733:Lepr APN 4 101622279 missense probably benign 0.00
IGL01815:Lepr APN 4 101671987 missense possibly damaging 0.49
IGL01899:Lepr APN 4 101637184 missense possibly damaging 0.86
IGL02138:Lepr APN 4 101625264 missense probably damaging 0.98
IGL02161:Lepr APN 4 101602875 missense probably damaging 0.97
IGL02653:Lepr APN 4 101622141 missense probably benign 0.44
IGL02735:Lepr APN 4 101639835 missense probably damaging 1.00
IGL03035:Lepr APN 4 101622177 missense probably damaging 1.00
IGL03083:Lepr APN 4 101671876 nonsense probably null
IGL03160:Lepr APN 4 101622103 missense probably damaging 1.00
aufsetzigen UTSW 4 101609372 missense probably damaging 1.00
beastly UTSW 4 101671788 missense probably benign
business_class UTSW 4 101622069 missense probably damaging 1.00
cherub UTSW 4 101625259 missense probably benign 0.25
clodhopper UTSW 4 101622487 splice site probably null
donner UTSW 4 101672398 missense probably damaging 1.00
fluffy UTSW 4 101649220 missense probably damaging 1.00
giant UTSW 4 101622349 critical splice donor site probably null
Immunoglutton UTSW 4 101622498 splice site probably benign
Jumbo_shrimp UTSW 4 101622151 nonsense probably null
lowleaning UTSW 4 101671588 splice site probably null
odd UTSW 4 101585271 splice site probably benign
paleo UTSW 4 101602842 missense possibly damaging 0.94
R0140_Lepr_245 UTSW 4 101625264 missense probably damaging 1.00
well-upholstered UTSW 4 101630155 synonymous probably benign
worldly UTSW 4 101625425 missense possibly damaging 0.96
PIT4651001:Lepr UTSW 4 101649194 missense probably damaging 1.00
PIT4696001:Lepr UTSW 4 101637180 missense probably benign 0.10
R0140:Lepr UTSW 4 101625264 missense probably damaging 1.00
R0197:Lepr UTSW 4 101609349 missense possibly damaging 0.64
R0279:Lepr UTSW 4 101607541 missense probably benign 0.05
R0487:Lepr UTSW 4 101625290 nonsense probably null
R0498:Lepr UTSW 4 101602889 missense probably benign 0.01
R0506:Lepr UTSW 4 101630207 splice site probably benign
R0512:Lepr UTSW 4 101671901 missense possibly damaging 0.87
R0512:Lepr UTSW 4 101649216 missense probably damaging 1.00
R0726:Lepr UTSW 4 101622131 missense probably benign 0.01
R1054:Lepr UTSW 4 101639793 missense probably damaging 0.97
R1109:Lepr UTSW 4 101628552 missense probably damaging 1.00
R1398:Lepr UTSW 4 101649216 missense probably damaging 1.00
R1464:Lepr UTSW 4 101592878 missense probably benign 0.08
R1464:Lepr UTSW 4 101592878 missense probably benign 0.08
R1519:Lepr UTSW 4 101646541 missense probably damaging 0.97
R1602:Lepr UTSW 4 101602842 missense possibly damaging 0.94
R1830:Lepr UTSW 4 101592874 missense probably damaging 1.00
R1850:Lepr UTSW 4 101590620 missense possibly damaging 0.67
R1918:Lepr UTSW 4 101630033 missense probably benign 0.08
R1928:Lepr UTSW 4 101639927 splice site probably benign
R2099:Lepr UTSW 4 101630185 missense probably damaging 1.00
R2102:Lepr UTSW 4 101630178 missense possibly damaging 0.95
R2175:Lepr UTSW 4 101622576 missense probably benign 0.01
R2254:Lepr UTSW 4 101672309 missense probably benign 0.26
R2396:Lepr UTSW 4 101590725 missense probably benign 0.19
R2508:Lepr UTSW 4 101648093 missense probably damaging 0.98
R2571:Lepr UTSW 4 101625369 missense possibly damaging 0.96
R3790:Lepr UTSW 4 101648111 splice site probably benign
R3882:Lepr UTSW 4 101672462 missense probably damaging 1.00
R3933:Lepr UTSW 4 101622498 splice site probably benign
R4211:Lepr UTSW 4 101590611 missense probably benign 0.19
R4343:Lepr UTSW 4 101622349 critical splice donor site probably null
R4345:Lepr UTSW 4 101622349 critical splice donor site probably null
R4544:Lepr UTSW 4 101625425 missense possibly damaging 0.96
R4546:Lepr UTSW 4 101671838 missense probably benign 0.35
R4724:Lepr UTSW 4 101622562 nonsense probably null
R4797:Lepr UTSW 4 101637244 missense possibly damaging 0.90
R4860:Lepr UTSW 4 101646534 missense probably benign 0.14
R4860:Lepr UTSW 4 101646534 missense probably benign 0.14
R4929:Lepr UTSW 4 101672314 missense probably benign 0.00
R4939:Lepr UTSW 4 101590635 missense possibly damaging 0.78
R5377:Lepr UTSW 4 101672216 missense possibly damaging 0.71
R5520:Lepr UTSW 4 101602734 missense probably benign 0.00
R5966:Lepr UTSW 4 101649324 intron probably benign
R6092:Lepr UTSW 4 101649220 missense probably damaging 1.00
R6130:Lepr UTSW 4 101622569 missense probably damaging 0.99
R6168:Lepr UTSW 4 101592789 missense probably damaging 0.99
R6232:Lepr UTSW 4 101671588 splice site probably null
R6380:Lepr UTSW 4 101622151 nonsense probably null
R6427:Lepr UTSW 4 101631454 missense possibly damaging 0.47
R6428:Lepr UTSW 4 101637295 missense probably damaging 1.00
R6641:Lepr UTSW 4 101622502 missense probably damaging 0.97
R6650:Lepr UTSW 4 101672398 missense probably damaging 1.00
R6859:Lepr UTSW 4 101622487 splice site probably null
R7023:Lepr UTSW 4 101646484 missense probably damaging 1.00
R7145:Lepr UTSW 4 101609394 missense probably benign 0.00
R7174:Lepr UTSW 4 101607535 missense probably benign 0.01
R7179:Lepr UTSW 4 101602856 missense probably benign 0.06
R7189:Lepr UTSW 4 101671961 missense probably benign 0.00
R7426:Lepr UTSW 4 101602853 missense probably benign 0.03
R7531:Lepr UTSW 4 101609372 missense probably damaging 1.00
R7620:Lepr UTSW 4 101609270 missense probably benign 0.41
R7804:Lepr UTSW 4 101639783 missense probably damaging 1.00
R8022:Lepr UTSW 4 101639754 missense probably benign 0.32
R8142:Lepr UTSW 4 101622616 missense possibly damaging 0.93
R8227:Lepr UTSW 4 101628559 missense probably damaging 0.99
R8426:Lepr UTSW 4 101671841 missense probably benign 0.12
R8447:Lepr UTSW 4 101671688 missense probably benign 0.08
R8531:Lepr UTSW 4 101622612 missense probably damaging 1.00
R8682:Lepr UTSW 4 101649269 missense probably benign 0.00
R8897:Lepr UTSW 4 101649233 missense probably damaging 0.98
R9096:Lepr UTSW 4 101631418 missense possibly damaging 0.95
R9177:Lepr UTSW 4 101602798 nonsense probably null
R9241:Lepr UTSW 4 101671788 missense probably benign
R9604:Lepr UTSW 4 101590473 missense probably benign 0.01
R9711:Lepr UTSW 4 101592851 nonsense probably null
X0026:Lepr UTSW 4 101590524 missense possibly damaging 0.47
Z1176:Lepr UTSW 4 101602811 missense probably damaging 0.99
Z1177:Lepr UTSW 4 101592792 missense probably damaging 1.00
Mode of Inheritance Unknown
Local Stock
Repository
Last Updated 2019-09-04 9:28 PM by Diantha La Vine
Record Created 2019-02-13 8:29 PM by Bruce Beutler
Record Posted 2019-02-18
Phenotypic Description

Figure 1. Gordo mice exhibit increased body weights. Scaled body weights are shown. Abbreviations: WT, wild-type; REF, homozygous reference mice; HET, heterozygous variant mice; VAR, homozygous variant mice. Mean (μ) and standard deviation (σ) are indicated.

The gordo phenotype was identified among N-Nitroso-N-ethylurea (ENU)-mutagenized G3 mice of the pedigree R6641, some of which showed increased body weights compared to wild-type littermates (Figure 1).

Nature of Mutation

Figure 2. Linkage mapping of the fat mass phenotype using a recessive model of inheritance. Manhattan plot shows -log10 P values (Y-axis) plotted against the chromosome positions of 36 mutations (X-axis) identified in the G1 male of pedigree R6641. Weight data are shown for single locus linkage analysis without consideration of G2 dam identity. Horizontal pink and red lines represent thresholds of P = 0.05, and the threshold for P = 0.05 after applying Bonferroni correction, respectively.

Whole exome HiSeq sequencing of the G1 grandsire identified 36 mutations. The body weight phenotype was linked to a mutation in Lepr: a T to A transversion at base pair 101,765,305 (v38) on chromosome 4, or base pair 48,018 in the GenBank genomic region NC_000070. Linkage was found with a recessive model of inheritance, wherein one variant homozygote departed phenotypically from 12 homozygous reference mice and 16 heterozygous mice with a P value of 6.891 x 10-7 (Figure 2).

The mutation corresponds to residue 1,832 in the mRNA sequence NM_146146 within exon 9 of 19 total exons.


 

1816 GAATTATACGTGATCGATGTCAATATCAATATA

422  -E--L--Y--V--I--D--V--N--I--N--I-

The mutated nucleotide is indicated in red. The mutation results in an aspartic acid to glutamic acid substitution at position 427 (D427E) in the LEPR protein, and is strongly predicted by PolyPhen-2 to be damaging (score = 0.972).

Illustration of Mutations in
Gene & Protein
Protein Prediction

Figure 3. Domain of OB-Rb and structure of mouse leptin receptor isoforms. OB-Rb contains the longest intracellular domain, which is crucial for leptin signaling. OB-Ra, OB-Rc and OB-Rd have varying short cytoplasmic domains. All isoforms contain the Box 1 motif known to bind JAK kinases. OB-Re is a secreted isoform of the leptin receptor. Cytokine receptor homology module (CRH)2 is the main binding site for leptin. The immunoglobin-like (IG-like) and fibronectin type III (FNIII) domains are involved in OB-R activation. The role of CRH1 remains to be determined. Both CRH domains also contain a FNIII fold (see text). The gordo mutation results in an aspartic acid to glutamic acid substitution at position 427 (D427E). This image is interactive. Other mutations found in the Lepr gene are noted in red. Click on the mutations for more specific information. 

The Lepr or obr gene encodes an 1162 amino acid protein that is the receptor for leptin, a four-helical cytokine-like hormone produced primarily by adipocytes [Figure 3; (1;2)]. The leptin receptor is a member of the gp130 family of cytokine receptors that are known to stimulate gene transcription via activation of cytosolic STAT (signal transducer and activator of transcription) proteins (see domino for more information on STAT signaling). Six isoforms of the leptin receptor are generated by alternative splicing; each isoform, with the exception of a soluble isoform (OB-Re) are single-pass membrane-spanning proteins differing only in the sequences of their C-terminal intracellular domains. The extracellular portion of the human leptin receptor contains two CK domains that contain conserved cysteine-containing motifs (amino acids 62-178 and amino acids 428-535), four domains that contain a fibronectin type III (FNIII) fold (amino acids 235-327, 536-635, 636-731, and amino acids 732-841), and a domain that has an Ig-like fold (amino acids 328-427). The first CK domain and the first FNIII domain form the cytokine receptor homology module 1 (CRH1), while the second CK domain and remaining FNIII domains form the cytokine receptor homology module 2. 

The gordo mutation results in an aspartic acid to glutamic acid substitution at position 427 (D427E); amino acid 427 is within the Ig-like domain.

Please see the record for Business_class for more information on Lepr.

Putative Mechanism

Leptin, a systemic hormone, regulates multiple functions of the body including energy utilization and storage, various endocrine axes, bone metabolism, thermoregulation, angiogenesis, immunity and inflammation by binding to the long form of the leptin receptor (OB-Rb) and subsequent initiation of various signal transduction pathways [reviewed in (3-5)].  It is primarily produced by adipocytes in proportion to fat stores, but can also be produced by placenta (syncytiotrophoblasts), ovaries, skeletal muscle, stomach, mammary epithelial cells, bone marrow, pituitary and liver (6). Humans and other animals deficient for leptin or its receptor, exhibit hyperphagia and low metabolism that results in obesity and insulin resistance [OMIM #614963; (2;7;8)].  Similar to other Lepr mouse models (see MGI for a list of Lepr alleles as well as the entry for Business_class), the gordo mice exhibit obesity. The phenotype of the gordo mice indicates that the LepRgordo protein exhibits loss of function.

Primers PCR Primer
gordo_pcr_F: AGGGAAGTTTACCTATGACGC
gordo_pcr_R: TTAACTGTGCCATCCCCAGG

Sequencing Primer
gordo_seq_F: CAGTGTACTGCTGCAATGAGC
gordo_seq_R: TCCCCAGGGAAACACAAAGAAG
Genotyping

PCR program

1) 94°C 2:00
2) 94°C 0:30
3) 55°C 0:30
4) 72°C 1:00
5) repeat steps (2-4) 40x
6) 72°C 10:00
7) 4°C hold


The following sequence of 400 nucleotides is amplified (chromosome 4, + strand):


1   agggaagttt acctatgacg cagtgtactg ctgcaatgag caggcgtgcc atcaccgcta
61  tgctgaatta tacgtgatcg gtaagaatcc caaggccgcg tccactccgc cctaggagat
121 catgtcatgg actgcgttag agtcctacta aagatgccag gaaaatcgtt gtattttttc
181 atgatttttg gtgtttttac tttatattaa tattttaatg tgttttaaat agatgtcaat
241 atcaatatat catgtgaaac tgacgggtac ttaactaaaa tgacttgcag atggtcaccc
301 agcacaatcc aatcactagt gggaagcact gtgcagctga ggtatcacag gtgtgtatgg
361 tactttcttc tttgtgtttc cctggggatg gcacagttaa 


Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red.

References
Science Writers Anne Murray
Illustrators Diantha La Vine
AuthorsZhao Zhang and Bruce Beutler