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|Mutation Type||critical splice donor site (1 bp from exon)|
|Coordinate||70,560,065 bp (GRCm38)|
|Base Change||G ⇒ A (forward strand)|
|Synonym(s)||ALUNC, AU, N, ba, bldy, hr, rh, rh-bmh, rhino|
|Chromosomal Location||70,552,212-70,573,548 bp (+)|
FUNCTION: This gene encodes a protein that is involved in hair growth. This protein functions as a transcriptional corepressor of multiple nuclear receptors, including thyroid hormone receptor, the retinoic acid receptor-related orphan receptors and the vitamin D receptors, and it interacts with histone deacetylases. The translation of this protein is modulated by a regulatory ORF that exists upstream of the primary ORF. Mutations in this upstream ORF, U2HR, cause Marie Unna hereditary hypotrichosis (MUHH), an autosomal dominant form of genetic hair loss in human. [provided by RefSeq, Oct 2014]
PHENOTYPE: Mutant homozygotes exhibit hair loss, usually wrinkled skin with epidermal cysts. Females do not nurse their pups well. [provided by MGI curators]
|Amino Acid Change|
|Institutional Source||Beutler Lab|
Ensembl: ENSMUSP00000022691 (fasta)
Ensembl: ENSMUSP00000124042 (fasta)
|Gene Model||not available|
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Recessive|
|Local Stock||Sperm, gDNA|
|Last Updated||2018-03-02 4:47 PM by Diantha La Vine|
The mister clean mutant phenotype emerged as a visible variant among G3 mice homozygous for mutations induced by N-ethyl-N-nitrosourea (ENU). The index mouse was completely hairless by weaning age and exhibited wrinkled skin (Figure 1). They are phenotypically similar to prune mice, which have a mutation in the hr gene.
|Nature of Mutation|
Due to the similarity in phenotype with prune mice, the hr locus was directly sequenced and a G to A transition was found in the donor splice site of intron 6 one nucleotide after the last exon (GTGAGC -> ATGAGC) in the hr gene on chromosome 14 (position 6009 in Genbank genomic region NC_000080 for linear genomic DNA sequence of hr). The mutation may result in skipping of the 191-nucleotide exon 6 (out of 20 total exons) in ENSMUST00000022691, destroying the reading frame in the middle of the encoded HR polypeptide chain (33 aberrant amino acids after position 515), and creating a premature stop codon that would truncate the protein after amino acid 548. The effect of the mutation at the cDNA and protein level has not been tested.
The donor splice site of intron 6, which is destroyed by the mister clean mutation, is indicated in blue lettering; the mutated nucleotide is indicated in red lettering.
In the ENSMUST00000163060 transcript, the mutation corresponds to the donor splice site of intron 4. The mutation may result in skipping of the 191-nucleotide exon 4 (out of 18 total exons), destroying the reading frame in the middle of the encoded HR polypeptide chain (33 aberrant amino acids after position 515), and creating a premature stop codon that would truncate the protein after amino acid 642.
The mister clean mutation likely results in the loss or weakening of the normal donor splice site of intron 4, which may lead to skipping of exon 4, a frame shift and premature truncation after the addition of 33 aberrant amino acids (Figure 2). Exon 4 does not encode any defined portion of the HR protein. Truncation occurs after the localization sequences and the first repressor domain. If the truncated protein was expressed, it would be missing the zinc finger, the JmjC domain, as well as two defined repression domains and nuclear receptor interacting motifs.
Please see the record for prune for more information about Hairless.
The phenotype of hr alleles is correlated with genotype. Thus, classical hairless mice that exhibit hairlessness, but not thickened and wrinkled skin, typically express some level of functional protein, while animals with severe versions of the rhino phenotype do not. The mister clean mutation is predicted to result in truncation of the HR protein, similar to some of the rhino alleles (1). Although mister clean mutants display some thickening and wrinkling of the skin, their phenotypes do not appear as severe as prune or knockout hr animals suggesting that some correct or alternative splicing may occur in these animals along with the retention of some HR function.
|Primers||Primers cannot be located by automatic search.|
Mister clean 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
MrClean(F): 5’- CCATCCGTCCACGTTGGAATCATC -3’
MrClean(R): 5’- TGCACTGGCTGTTTCCTCCATAAG -3’
PCR program (use SIGMA JumpStart REDTaq)
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
MrClean_seq(F1): 5’- CTTCCTAGAGACTTGGGCTGAAC -3'
MrClean_seq(F2): 5’- AGACTTGTGGATACATCCTGC -3’
MrClean_seq(R): 5’- GACTGGACCAAGTCTTTAGACTC -3’
The following sequence of 1033 nucleotides (from Genbank genomic region NC_000080 for linear DNA sequence of hr) is amplified:
5210 c catccgtcca
5221 cgttggaatc atctcacagt gtcacatagc cacttagcct ttagctgacc tacccaaagt
5281 ctatactggc tggattctgc acgtgtacaa ggaccattgc tcaggcttcc tagagacttg
5341 ggctgaacat gctaaacaac ccctgagcct ccattcctac cagggacatc tccagctact
5401 gctcagcttt gtgtgtgaaa tatactggac tgatcctctt tcgggttctg aattggtcct
5461 cacaagcctg gggtccaggg gctgaattgg ctgcatgtgg acaagggtgg gtgttaattc
5521 agggctttga ctagcataag ccccagaaac cagactccct aagcaactgt attgtctccc
5581 cagggccccg agatggcagg attaggctcc aggagtccag acttgtggat acatcctgcc
5641 agcatcactt agcaggtgtc acccagtgcc aaagctgtgt ccaggcagct ggagaggtag
5701 gggtactgac cggccactcc cagaaatcac gtaggtgagt gttgtgtctg acagtcagag
5761 ccagcagcac tccatcccca cccaggggct ccctcctcaa gcttcatcca tttccaggtc
5821 acccctggag gagaagcagt tggaggagga ggattcctct gccacttccg aagaaggagg
5881 aggagggcct ggcccagaag cttcactcaa caagggcctg gccaagcacc tgctgagtgg
5941 tttgggggac cgactctgcc gcctgctgcg gaaggagcgg gaggcccttg cctgggcaca
6001 gcgagaaggt gagccaattt cccttgtggg cctgctcctg catgtccctc ccaacccacg
6061 tttaccagtg ctttggggtt cccaggccta ctcaggagag ctccgtcctt ctttgttcct
6121 agctatccct agcacggact cttggtcaat cctgaggttt ctggcctcct ggggagtcta
6181 aagacttggt ccagtctagt tctctaagaa caactgggct tatggaggaa acagccagtg
PCR primer binding sites are underlined; sequencing primer binding sites are highlighted in gray; the mutated G is shown in red text.
1. Cachon-Gonzalez, M. B., San-Jose, I., Cano, A., Vega, J. A., Garcia, N., Freeman, T., Schimmang, T., and Stoye, J. P. (1999) The hairless gene of the mouse: relationship of phenotypic effects with expression profile and genotype, Dev. Dyn. 216, 113-126.
|Science Writers||Nora G. Smart|
|Illustrators||Diantha La Vine|
|Authors||Xin Du, Bruce Beutler|
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