Phenotypic Mutation 'giant' (pdf version)
Allele | giant |
Mutation Type |
critical splice donor site
|
Chromosome | 4 |
Coordinate | 101,622,349 bp (GRCm39) |
Base Change | G ⇒ 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
|
Mapped | Yes |
Limits of the Critical Region |
101717404 - 101815352 bp |
Amino Acid Change |
|
Institutional Source | Beutler Lab |
Gene Model |
predicted gene model for protein(s):
[ENSMUSP00000037385 †]
[ENSMUSP00000099838 †]
[ENSMUSP00000102534 †]
† probably from a misspliced transcript
|
AlphaFold |
P48356 |
SMART Domains |
Protein: ENSMUSP00000037385 Gene: ENSMUSG00000057722
Domain | Start | End | E-Value | Type |
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 null
|
SMART Domains |
Protein: ENSMUSP00000099838 Gene: ENSMUSG00000057722
Domain | Start | End | E-Value | Type |
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 null
|
SMART Domains |
Protein: ENSMUSP00000102534 Gene: ENSMUSG00000057722
Domain | Start | End | E-Value | Type |
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 null
|
Meta Mutation Damage Score |
0.9484 |
Is this an essential gene? |
Non Essential (E-score: 0.000) |
Phenotypic Category |
Autosomal Recessive |
Candidate Explorer Status |
loading ... |
Single pedigree Linkage Analysis Data
|
|
Penetrance | |
Alleles Listed at MGI | All Mutations and Alleles(53) : Chemically induced (ENU)(9) Chemically induced (other)(2) Radiation induced(1) Spontaneous(15) Targeted(23) Transgenic(3)
|
Lab Alleles |
Allele | Source | Chr | Coord | Type | Predicted Effect | PPH 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 |
gordo
|
UTSW |
4 |
101622502 |
missense |
probably damaging |
0.97 |
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 |
Autosomal Recessive |
Local Stock | |
Repository | |
Last Updated |
2019-09-04 9:42 PM
by Diantha La Vine
|
Record Created |
2016-07-27 6:52 PM
by Jin Huk Choi
|
Record Posted |
2016-09-15 |
Phenotypic Description |
The giant phenotype was identified among N-Nitroso-N-ethylurea (ENU)-mutagenized G3 mice of the pedigree R4345, some of which showed increased body weights (Figure 1 & 2), increased blood glucose levels 30 minutes after glucose bolus (Figure 3), increased levels of IgE levels in the peripheral blood (Figure 4), and reduced killing of natural killer target cells (Figure 5) compared to wild-type mice.
|
Nature of Mutation |
Whole exome HiSeq sequencing of the G1 grandsire identified 41 mutations. All of the above anomalies were linked by continuous variable mapping to a mutation in Lepr: a G to A transition at base pair 101,765,152 (v38) on chromosome 4, or base pair 48,019 in the GenBank genomic region NC_000070 within the donor splice site on intron 8 of the Lepr gene. The strongest association was found with a recessive model of linkage to the body weight phenotype, wherein two variant homozygotes departed phenotypically from six homozygous reference mice and 15 heterozygous mice with a P value of 1.213 x 10-17 (Figure 6). The effect of the mutation at the cDNA and protein level have not examined, but the mutation is predicted to result in skipping of the 291-nucleotide exon 8 (out of 19 total exons), resulting in an in-frame deletion of 97 amino acids beginning after amino acid 329 of the encoded protein.
<--exon 7 <--exon 8 intron 8--> exon 9--> <--exon 19
35057 ……GTCTTTACCACACAAG ……GAATTATACGTGATCG gtaagaatcccaagg…… ATGTCAATATCAAT…… ……GACTTAACTGTGTAA 98122
325 ……-V--F--T--T--Q-- ……-E--L--Y--V--I-- D--V--N--I--N-…… ……-D--L--T--V--*- 1162
correct deleted correct
|
The donor splice site of intron 8, which is destroyed by the giant mutation, is indicated in blue lettering and the mutated nucleotide is indicated in red.
|
Illustration of Mutations in
Gene & Protein |
|
---|
Protein Prediction |
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 7; (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 giant mutation is a donor splice site mutation in intron 8 that is predicted to result in the in-frame deletion of exon 8. Exon 8 encodes amino acids 330-426, which encompasses the Ig-like fold 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 giant mice exhibit obesity. The phenotype of the giant mice indicates that the LepRgiant protein exhibits loss of function.
|
Primers |
PCR Primer
giant_pcr_F: AGGAATCTAGCTGAGAAAATCCCTG
giant_pcr_R: GTGCTTCCCACTAGTGATTGG
Sequencing Primer
giant_seq_F: CCCTGAGATACAGTACAGCATTGTG
giant_seq_R: AAGTACCCGTCAGTTTCACATG
|
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 427 nucleotides is amplified (chromosome 4, + strand):
1 aggaatctag ctgagaaaat ccctgagata cagtacagca ttgtgagtga ccgagttagc 61 aaagttacct tctccaacct gaaagccacc agacctcgag ggaagtttac ctatgacgca 121 gtgtactgct gcaatgagca ggcgtgccat caccgctatg ctgaattata cgtgatcggt 181 aagaatccca aggccgcgtc cactccgccc taggagatca tgtcatggac tgcgttagag 241 tcctactaaa gatgccagga aaatcgttgt attttttcat gatttttggt gtttttactt 301 tatattaata ttttaatgtg ttttaaatag atgtcaatat caatatatca tgtgaaactg 361 acgggtactt aactaaaatg acttgcagat ggtcacccag cacaatccaa tcactagtgg 421 gaagcac
Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red. |
References | 1. Zhang, F., Basinski, M. B., Beals, J. M., Briggs, S. L., Churgay, L. M., Clawson, D. K., DiMarchi, R. D., Furman, T. C., Hale, J. E., Hsiung, H. M., Schoner, B. E., Smith, D. P., Zhang, X. Y., Wery, J. P., and Schevitz, R. W. (1997) Crystal Structure of the Obese Protein Leptin-E100. Nature. 387, 206-209.
2. Zhang, Y., Proenca, R., Maffei, M., Barone, M., Leopold, L., and Friedman, J. M. (1994) Positional Cloning of the Mouse Obese Gene and its Human Homologue. Nature. 372, 425-432.
4. Malendowicz, L. K., Rucinski, M., Belloni, A. S., Ziolkowska, A., and Nussdorfer, G. G. (2007) Leptin and the Regulation of the Hypothalamic-Pituitary-Adrenal Axis. Int Rev Cytol. 263, 63-102.
7. Chen, H., Charlat, O., Tartaglia, L. A., Woolf, E. A., Weng, X., Ellis, S. J., Lakey, N. D., Culpepper, J., Moore, K. J., Breitbart, R. E., Duyk, G. M., Tepper, R. I., and Morgenstern, J. P. (1996) Evidence that the Diabetes Gene Encodes the Leptin Receptor: Identification of a Mutation in the Leptin Receptor Gene in db/db Mice. Cell. 84, 491-495.
8. Farooqi, I. S., Wangensteen, T., Collins, S., Kimber, W., Matarese, G., Keogh, J. M., Lank, E., Bottomley, B., Lopez-Fernandez, J., Ferraz-Amaro, I., Dattani, M. T., Ercan, O., Myhre, A. G., Retterstol, L., Stanhope, R., Edge, J. A., McKenzie, S., Lessan, N., Ghodsi, M., De, R.,V, Perna, F., Fontana, S., Barroso, I., Undlien, D. E., and O'Rahilly, S. (2007) Clinical and Molecular Genetic Spectrum of Congenital Deficiency of the Leptin Receptor. N Engl J Med. 356, 237-247.
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Science Writers | Anne Murray |
Illustrators | Peter Jurek |
Authors | Jin Huk Choi, Tao Yue, Evan Nair-Gill, and Bruce Beutler |