Phenotypic Mutation 'rollo' (pdf version)
Allelerollo
Mutation Type nonsense
Chromosome1
Coordinate52,193,082 bp (GRCm39)
Base Change C ⇒ A (forward strand)
Gene Stat1
Gene Name signal transducer and activator of transcription 1
Synonym(s) 2010005J02Rik
Chromosomal Location 52,158,599-52,201,024 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 is a member of the STAT protein family. In response to cytokines and growth factors, STAT family members are phosphorylated by the receptor associated kinases, and then form homo- or heterodimers that translocate to the cell nucleus where they act as transcription activators. This protein can be activated by various ligands including interferon-alpha, interferon-gamma, EGF, PDGF and IL6. This protein mediates the expression of a variety of genes, which is thought to be important for cell viability in response to different cell stimuli and pathogens. Two alternatively spliced transcript variants encoding distinct isoforms have been described. [provided by RefSeq, Jul 2008]
PHENOTYPE: Homozygotes for targeted null mutations are largely unresponsive to interferon, fail to thrive, are susceptible to viral diseases and cutaneous leishmaniasis, and show excess osteoclastogenesis leading to increased bone mass. [provided by MGI curators]
Accession Number

NCBI RefSeq: NM_001205313 (variant 1), NM_009283 (variant 2), NM_001205314 (variant 3); MGI:103063

MappedYes 
Limits of the Critical Region 52119440 - 52161865 bp
Amino Acid Change Tyrosine changed to Stop codon
Institutional SourceBeutler Lab
Gene Model predicted gene model for protein(s): [ENSMUSP00000066743] [ENSMUSP00000141132] [ENSMUSP00000140518] [ENSMUSP00000140875] [ENSMUSP00000140482] [ENSMUSP00000141125] [ENSMUSP00000139746]
AlphaFold no structure available at present
SMART Domains Protein: ENSMUSP00000066743
Gene: ENSMUSG00000026104
AA Change: Y651*

DomainStartEndE-ValueType
STAT_int 2 122 2.5e-61 SMART
Pfam:STAT_alpha 139 315 1.4e-56 PFAM
Pfam:STAT_bind 317 566 4.2e-82 PFAM
SH2 571 687 1.59e-1 SMART
Pfam:STAT1_TAZ2bind 715 739 2.4e-17 PFAM
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000141132
Gene: ENSMUSG00000026104
AA Change: Y657*

DomainStartEndE-ValueType
STAT_int 2 122 2.5e-61 SMART
Pfam:STAT_alpha 136 315 3.4e-65 PFAM
Pfam:STAT_bind 317 573 3.9e-118 PFAM
SH2 577 693 1.59e-1 SMART
Pfam:STAT1_TAZ2bind 721 745 2.3e-16 PFAM
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000140518
Gene: ENSMUSG00000026104
AA Change: Y651*

DomainStartEndE-ValueType
STAT_int 2 122 1.9e-65 SMART
Pfam:STAT_alpha 136 315 3.3e-62 PFAM
Pfam:STAT_bind 317 567 1.1e-118 PFAM
SH2 571 687 1e-3 SMART
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000140875
Gene: ENSMUSG00000026104
AA Change: Y651*

DomainStartEndE-ValueType
STAT_int 2 122 2.5e-61 SMART
Pfam:STAT_alpha 136 315 1.2e-64 PFAM
Pfam:STAT_bind 317 567 4.4e-121 PFAM
SH2 571 687 1.59e-1 SMART
Pfam:STAT1_TAZ2bind 715 739 3.1e-15 PFAM
Predicted Effect probably null
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000140482
Gene: ENSMUSG00000026104
AA Change: Y145*

DomainStartEndE-ValueType
Pfam:STAT_bind 13 61 2.6e-14 PFAM
SH2 65 181 1.59e-1 SMART
Pfam:STAT1_TAZ2bind 209 233 7.8e-16 PFAM
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000141125
Gene: ENSMUSG00000026104
AA Change: Y651*

DomainStartEndE-ValueType
STAT_int 2 122 1.9e-65 SMART
Pfam:STAT_alpha 136 315 3.3e-62 PFAM
Pfam:STAT_bind 317 567 1.1e-118 PFAM
SH2 571 687 1e-3 SMART
Predicted Effect probably null
SMART Domains Protein: ENSMUSP00000139746
Gene: ENSMUSG00000026104
AA Change: Y651*

DomainStartEndE-ValueType
STAT_int 2 122 1.9e-65 SMART
Pfam:STAT_alpha 136 315 3.3e-62 PFAM
Pfam:STAT_bind 317 567 1.1e-118 PFAM
SH2 571 687 1e-3 SMART
Predicted Effect probably null
Meta Mutation Damage Score 0.9755 question?
Is this an essential gene? Non Essential (E-score: 0.000) question?
Phenotypic Category Autosomal Recessive
Candidate Explorer Status loading ...
Single pedigree
Linkage Analysis Data
Penetrance  
Alleles Listed at MGI

All Mutations and Alleles(23) : Chemically induced (ENU)(4) Chemically induced (other)(1) Radiation induced(1) Targeted(17)

Lab Alleles
AlleleSourceChrCoordTypePredicted EffectPPH Score
IGL00092:Stat1 APN 1 52161754 start codon destroyed probably null 0.50
IGL01111:Stat1 APN 1 52182120 critical splice donor site probably null
IGL01451:Stat1 APN 1 52178502 missense probably damaging 1.00
IGL01469:Stat1 APN 1 52186529 missense possibly damaging 0.87
IGL01758:Stat1 APN 1 52176080 missense probably damaging 1.00
IGL01818:Stat1 APN 1 52190437 missense probably damaging 1.00
IGL01913:Stat1 APN 1 52165716 missense probably benign 0.08
IGL01914:Stat1 APN 1 52165716 missense probably benign 0.08
IGL02304:Stat1 APN 1 52171703 missense probably benign
IGL02428:Stat1 APN 1 52182125 splice site probably benign
Accretion UTSW 1 52174780 missense possibly damaging 0.65
Aspect UTSW 1 52190408 missense probably benign 0.01
baroque UTSW 1 52183368 missense probably damaging 1.00
Compounding UTSW 1 52190440 missense probably benign 0.17
domino UTSW 1 52179747 missense probably damaging 1.00
h_moll UTSW 1 52178353 nonsense probably null
kun_ming UTSW 1 52176575 missense possibly damaging 0.52
kuomintang UTSW 1 52190404 missense possibly damaging 0.51
poison UTSW 1 52190384 splice site probably benign
roccoco UTSW 1 52162368 missense probably damaging 1.00
Sedimentary UTSW 1 52178388 missense probably damaging 1.00
special UTSW 1 52178423 missense probably damaging 1.00
vandegraff UTSW 1 52194178 missense probably benign 0.01
R0022:Stat1 UTSW 1 52179789 missense probably damaging 1.00
R0022:Stat1 UTSW 1 52179789 missense probably damaging 1.00
R0039:Stat1 UTSW 1 52179819 missense probably damaging 0.99
R0458:Stat1 UTSW 1 52188211 splice site probably benign
R1313:Stat1 UTSW 1 52195165 missense probably damaging 0.98
R1313:Stat1 UTSW 1 52195165 missense probably damaging 0.98
R2998:Stat1 UTSW 1 52190408 missense probably benign 0.01
R4464:Stat1 UTSW 1 52176575 missense possibly damaging 0.52
R4709:Stat1 UTSW 1 52165680 missense probably damaging 0.97
R4934:Stat1 UTSW 1 52193082 nonsense probably null
R5038:Stat1 UTSW 1 52162368 missense probably damaging 1.00
R5075:Stat1 UTSW 1 52161871 missense possibly damaging 0.73
R5223:Stat1 UTSW 1 52183401 missense probably damaging 1.00
R5600:Stat1 UTSW 1 52188101 missense probably benign 0.06
R5866:Stat1 UTSW 1 52178423 missense probably damaging 1.00
R7105:Stat1 UTSW 1 52190408 missense probably benign 0.01
R7192:Stat1 UTSW 1 52174780 missense possibly damaging 0.65
R7284:Stat1 UTSW 1 52188081 missense probably benign 0.01
R7309:Stat1 UTSW 1 52165780 splice site probably null
R7491:Stat1 UTSW 1 52191530 missense probably benign 0.31
R7680:Stat1 UTSW 1 52183368 missense probably damaging 1.00
R7825:Stat1 UTSW 1 52190467 missense probably damaging 0.98
R7915:Stat1 UTSW 1 52190440 missense probably benign 0.17
R8245:Stat1 UTSW 1 52194178 missense probably benign 0.01
R8309:Stat1 UTSW 1 52190404 missense possibly damaging 0.51
R8728:Stat1 UTSW 1 52178353 nonsense probably null
R8952:Stat1 UTSW 1 52187042 missense probably benign 0.01
R9054:Stat1 UTSW 1 52182086 missense probably damaging 1.00
R9156:Stat1 UTSW 1 52178388 missense probably damaging 1.00
R9209:Stat1 UTSW 1 52184337 missense probably benign
R9252:Stat1 UTSW 1 52174831 missense probably benign 0.03
R9337:Stat1 UTSW 1 52191429 missense probably benign 0.00
R9388:Stat1 UTSW 1 52193037 missense possibly damaging 0.81
R9530:Stat1 UTSW 1 52187160 critical splice donor site probably null
R9648:Stat1 UTSW 1 52165695 missense probably damaging 0.98
RF036:Stat1 UTSW 1 52191419 missense probably benign
RF060:Stat1 UTSW 1 52191419 missense probably benign
X0027:Stat1 UTSW 1 52178430 missense probably damaging 1.00
Mode of Inheritance Autosomal Recessive
Local Stock
Repository
Last Updated 2019-09-04 9:41 PM by Diantha La Vine
Record Created 2016-12-30 2:27 PM by Evan Nair-Gill
Record Posted 2017-09-15
Phenotypic Description

Figure 1. Rollo mice exhibited decreased ability to kill beta-2-microglobulin deficient cells (NK cell targets) compared to heterozygous or reference mice. Normalized data 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 rollo phenotype was identified among N-ethyl-N-nitrosourea (ENU)-mutagenized G3 mice of the pedigree R4934 some of which showed reduced ability to kill beta-2-microglobulin deficient cells (NK cell targets) compared to heterozygous or reference mice (Figure 1).

Nature of Mutation

Figure 2. Linkage mapping of the reduced NK cell target killing using a recessive model of inheritance. Manhattan plot shows -log10 P values (Y-axis) plotted against the chromosome positions of 132 mutations (X-axis) identified in the G1 male of pedigree R4934. Normalized phenotype 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 132 mutations. The NK killing phenotype was linked by continuous variable mapping to two mutations on chromosome 1: Col3a1 and Stat1. The mutation in Stat1 was presumed causative, and is a C to A transversion at base pair 52,153,923 (v38) on chromosome 1, or base pair 34,486 in the GenBank genomic region NC_000067 encoding Stat1. Linkage was found with a recessive model of inheritance, wherein four variant homozygotes departed phenotypically from nine homozygous reference mice and six heterozygous mice with a P value of 8.137 x 10-10 (Figure 2).  

The mutation corresponds to residue 2,270 in the mRNA sequence NM_001205313 within exon 22 of 25 total exons.

2253 GATATTATTCGCAACTACAAAGTCATGGCTGCC

646  -D--I--I--R--N--Y--K--V--M--A--A-

The mutated nucleotide is indicated in red. The mutation results in substitution of tyrosine 651 for a premature stop codon (Y651*) in the STAT1 protein (Figure 3).

Illustration of Mutations in
Gene & Protein
Protein Prediction
Figure 3. 3D and domain structure of the STAT1 protein. A) 3D representation of STAT1 based on crystalized structures of human STAT1 residues 1-683 (PDB 1YVL). The residue affected by the domino mutation is shown in red. 3D image was created using UCSF Chimera. B) Domain structure of STAT1. CC=Coiled Coil domain; DBD = DNA binding domain; LD = Linker domain; SH2=Src Homology 2 domain; TAD = Transcriptional activation domain. The critical tyrosine phosphorylation site is found at amino acid 701. The rollo mutation substitution of tyrosine 651 for a premature stop codon (Y651*) . This image is interactive. Other mutations found in STAT1 are noted in red. Click on the mutations for more specific information. Click on the 3D structure to view it rotate. 

Signal transducer and activator of transcription (STAT)-1 is one of seven STAT family members identified in mammals. The STAT proteins serve the dual functions of signal transduction and activation of transcription. STAT1 is a 755 amino acid protein, and like all STATs, contains an N-terminal helical domain (N-domain), a four helix bundle, a central Ig-like DNA binding domain, a helical linker domain, an SH2 domain, and a C-terminal transactivation domain (TAD) (Figure 3).

The rollo mutation results in substitution of tyrosine 651 for a premature stop codon (Y651*) in the STAT1 protein; amino acid 651 is within the SH2 domain.

Please see the record for domino for more information about Stat1.

Putative Mechanism

The STAT proteins are transcription factors found latent in the cytoplasm until they are activated by extracellular signaling proteins such as cytokines, growth factors and peptides. Stimulation by these extracellular signaling proteins leads to activation of intracellular tyrosine kinases that in turn phosphorylate STATs, causing them to move into the nucleus and activate transcription of target genes. STAT1 is required for IFN signaling.  NK cell function is enhanced by IFNs, and Stat1-/- NK cells have impaired cytotoxicity relative to wild type NK cells (1)Stat1-/- mice have no gross developmental abnormalities, but are highly sensitive to bacterial and viral infections such as Listeria monocytogenes and VSV infection (2;3). Cells from these mice are unresponsive to IFN-α and IFN-γ, although they respond normally to several other stimuli including EGF and interleukin 10 (2;3). In humans, rare STAT1 deficiency and several STAT1 point mutations have been identified in patients with recurrent bacterial and/or viral infections (4-6). Cells from these patients fail to respond to IFN-α or IFN-γ. Interestingly, one patient with complete STAT1 deficiency was able to clear at least some viruses including polio virus type III (from vaccination) and parainfluenza type II (6).

The phenotype observed in the rollo mice indicates loss of STAT1rollo function.

Primers PCR Primer
rollo_pcr_F: TTAAAGCAGGAGTGTGATGCC
rollo_pcr_R: AACTTGTCTGGGAGCCATGG

Sequencing Primer
rollo_seq_F: AAAGTATGGCAGGCTCTCTG
rollo_seq_R: TCTGGGAGCCATGGAGCAC
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 401 nucleotides is amplified (chromosome 1, + strand):


1   ttaaagcagg agtgtgatgc ctttaaaaaa gtatggcagg ctctctgact ccccctgggc
61  tgtgtctgcc ttgctttagt ggtctctggg gcgcctggga ctcactggac ctcactggtt
121 tcctccttgt agaacctgac ttccatgccg tggagcccta cacgaaaaaa gaactttcag
181 ctgttacttt cccagatatt attcgcaact acaaagtcat ggctgccgag aacataccag
241 agaatcccct gaagtatctg taccccaata ttgacaaaga ccacgccttt gggaagtatt
301 attccagacc aaaggaaggt gagtgcctga gcgatctggg gtgcctgtgg gggagagtgc
361 agcaaggccc ttcctggtgc tccatggctc ccagacaagt t


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, Peter Jurek, Katherine Timer
AuthorsEvan Nair-Gill, Bruce Beutler