|Mutation Type||splice site (11 bp from exon)|
|Coordinate||34,258,184 bp (GRCm38)|
|Base Change||A ⇒ T (forward strand)|
|Gene Name||dedicator of cyto-kinesis 2|
|Synonym(s)||CED-5, MBC, Hch|
|Chromosomal Location||34,226,815-34,783,892 bp (-)|
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 CDM protein family. It is specifically expressed in hematopoietic cells and is predominantly expressed in peripheral blood leukocytes. The protein is involved in remodeling of the actin cytoskeleton required for lymphocyte migration in response to chemokine signaling. It activates members of the Rho family of GTPases, for example RAC1 and RAC2, by acting as a guanine nucleotide exchange factor (GEF) to exchange bound GDP for free GTP. [provided by RefSeq, Oct 2016]
PHENOTYPE: Homozygous mutants are defective in the migration of T and B lympohcytes in response to chemokines, and thus display immune defects such as lymphocytopenia, atrophy of lymphoid follicles and loss of marginal-zone B cells. [provided by MGI curators]
|Amino Acid Change|
|Institutional Source||Beutler Lab|
|Gene Model||not available|
|Predicted Effect||probably benign|
|Meta Mutation Damage Score||Not available|
|Is this an essential gene?||Non Essential (E-score: 0.000)|
|Candidate Explorer Status||CE: no linkage results|
Linkage Analysis Data
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Recessive|
|Local Stock||Sperm, gDNA|
|Last Updated||2019-05-21 7:37 PM by Diantha La Vine|
|Record Created||2010-03-15 3:55 PM by Carrie N. Arnold|
|Other Mutations in This Stock||
Stock #: G5538 Run Code: SLD00239
Coding Region Coverage: 1x: 75.9% 3x: 46.5%
Validation Efficiency: 69/84
The frizz mutation was discovered while analyzing N-ethyl-N-nitrosourea (ENU)-mutagenized G3 mice using flow cytometry analysis of blood. Frizz animals have reduced frequencies of peripheral CD4+ and CD8+ T cells and low expression of B220 (the B cell form of CD45; see the record for belittle), suggesting a block in B cell maturation (Figure 1). In addition, these animals lack a T-dependent immunoglobin (IgG) response to model antigens encoded by a recombinant nonreplicating vector based on the Semliki Forest Virus (rSFV), but make normal T-independent IgM responses to haptenated ficoll (Figure 2). The frizz phenotype is similar to that of frazz mutants with a mutation in the Dock2 gene. Complementation testing between the two strains confirmed that frizz is also an allele of Dock2.
|Nature of Mutation|
Whole genome sequencing of a homozygous frizz mouse using the SOLiD technique covered the coding/splicing region at least 1x or 3x with 75.9% and 46.5% coverage, respectively. Validation sequencing using the Sanger method was attempted on all nucleotides for which discrepancies were seen at 3x or greater coverage, with 69 of 84 discrepancies successfully processed and mutations in three genes were identified. Whole genome sequencing did not identify the causative mutation in Dock2. Standard Sanger sequencing of the Dock2 gene identified a T to A transversion at position 34158184 in the GenBank genomic region NC_000077 for the Dock2 gene on chromosome 11 (TGATCCCATAG -> AGATCCCATAG) (Figure). In GenBank, the mutation is located near the acceptor splice site of intron 16 (intron 38 in Ensembl record ENSMUST00000093193) from the ATG exon, eleven nucleotides to the next exon. Dock2 contains 30 exons according to Genbank record NM_033374 and 52 exons according to Ensembl record ENSMUST00000093193. Multiple Dock2 transcripts are displayed on Ensembl and Vega. The effect of the mutation at the cDNA and protein level is currently being determined. One possibility, shown below, is that aberrant splicing may result in skipping of the 90 bp exon 39, utilization of the acceptor splice site from intron 39 and in-frame splicing to exon 40 (depicted below as seen on Ensembl). This would result in deletion of 30 amino acids.
<--exon 38 <--intron 38 exon 39--> exon 40--> <--exon 52
1289 -K--G--K- -M--W--E- -T--Q--Q-………-N--M--* 1828
correct deleted correct
The frizz mutation likely results in abnormal splicing of Dock2 and may cause an internal deletion of amino acids 1292-1321 in the DHR-2 domain.
For more information on the Dock2 gene, see the record for frazz.
|Primers||Primers cannot be located by automatic search.|
Frizz genotyping is performed by amplifying the region containing the mutation using PCR, followed by sequencing of the amplified region to detect the single nucleotide transition.
Frizz(F): 5’- GATGCTCACCAAGACCTCATCAGG -3’
Frizz(R): 5’- GCGCAGATGACTCTCCATTTCTCAC -3’
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 8
Primers for sequencing
Frizz_seq(F): 5'- AGACCTCATCAGGCCCTTC -3'
Frizz_seq(R): 5'- TCTCTCCCAGTACTTAGGAAGATCAG -3'
The following sequence of 513 nucleotides (NCBI Mouse Genome Build 37.1, Chromosome 11, bases 34,157,954 to 34,158,466) is amplified:
gcgcagatga ctctccattt ctcacctaga gtgtaggtct gatgttagtg acctctctcc cagtacttag aagatcaga gcacatgtaa acaagttact ccattattca gtaaaggagc tggttagaaa gccaggggaa gtttgtacta ctaccaaggc ccgctctgtg tgagcactag gtaagaagct gtctgcggtg caacgggcga tggcaaggct caagtccctc cttcagcaaa ggctctgaaa agttggacca cctctaacca ctgtccccat gatgatccca tagatgtggg aagaggccat cagcctgtgc aaggaactgg cggaacaata tgagatggag atctttgact acgagctact cagccagaac ctggtaaggc tctcccaaga aagccatgca cgccacacac gcggtgctcg ctcagctcag agacagaagc gaggcccgtg ggttagggag acagaagact ggggaagggc ctgatgaggt cttggtgagc atc
Primer binding sites are underlined; sequencing primer binding sites are highlighted in gray; the mutated T is indicated in red.
|Science Writers||Nora G. Smart|
|Authors||Carrie N. Arnold, Elaine Pirie, and Bruce Beutler|