|Coordinate||36,781,412 bp (GRCm38)|
|Base Change||T ⇒ C (forward strand)|
|Gene Name||zeta-chain (TCR) associated protein kinase|
|Synonym(s)||ZAP-70, TZK, Srk|
|Chromosomal Location||36,761,798-36,782,818 bp (+)|
|MGI Phenotype||Mutant mice show T cell defects. Null mutants lack alpha-beta T cells in the thymus and have fewer T cells in dendritic and intestinal epithelium. Spontaneous and knock-in missense mutations affect T cell receptor signaling, one of the former resulting in severe chronic arthritis.|
|Limits of the Critical Region||36761861 - 36782816 bp|
|Amino Acid Change||Cysteine changed to Arginine|
|Institutional Source||Beutler Lab|
|Gene Model||predicted gene model for protein(s): [ENSMUSP00000027291]|
AA Change: C563R
|Predicted Effect||probably damaging
PolyPhen 2 Score 1.000 (Sensitivity: 0.00; Specificity: 1.00)
|Phenotypic Category||Autosomal Recessive|
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Recessive|
|Last Updated||2017-08-08 8:10 PM by Diantha La Vine|
|Record Created||2015-09-18 10:31 PM by Jin Huk Choi|
The wanna2 phenotype was identified among G3 mice of the pedigree R3689, one of which showed a diminished T-dependent antibody response to recombinant Semliki Forest virus (rSFV)-encoded β-galactosidase (rSFV-β-gal) (Figure 1).
|Nature of Mutation|
Whole exome HiSeq sequencing of the G1 grandsire identified 57 mutations. The diminished T-dependent antibody response to rSFV-β-gal phenotype was linked by continuous variable mapping to a mutation in Zap70: a T to C transition at base pair 36,781,412 (v38) on chromosome 1, or base pair 19,615 in the GenBank genomic region NC_000067. Linkage was found with a recessive model of inheritance (P = 7.764 x 10-6), wherein one variant homozygote departed phenotypically from three homozygous reference mice and 10 heterozygous mice (Figure 2).
The mutation corresponds to residue 1,849 in the mRNA sequence NM_001289766 within exon 12 of 13 total exons.
Genomic numbering corresponds to NC_000067. The mutated nucleotide is indicated in red. The mutation results in a cysteine (C) to arginine (R) substitution at residue 563 (C563R) in the ZAP70 protein, and is strongly predicted by PolyPhen-2 to be damaging (score = 0.999) (1).
The ζ-associated protein of 70 kDa (ZAP-70) is a protein tyrosine kinase (PTK) that binds to the doubly phosphorylated immunoreceptor tyrosine-based activation motifs (ITAMS) of ζ and CD3ε chains of the T cell receptor (TCR; see the record for tumormouse). ZAP70 consists of two N-terminal Src-homology 2 (SH2) domains at amino acids and a C-terminal kinase domain. The SH2 domains are connected by a linker known as interdomain A, while the region between the second SH2 and catalytic domains is known as interdomain B (2). The two SH2 domains of mouse ZAP-70 occur at amino acids 10-102 and 163-254, and work cooperatively to bind to the phosphorylated tyrosines of an ITAM sequence [(D/E)xxYxxI/Lx(6-8)YxxI/L]. The wanna2 mutation results in a cysteine (C) to arginine (R) substitution at 563 within the ZAP70 kinase domain (Figure 3).
Please see the record for murdock for more information about Zap70.
Signaling through the T cell receptor (TCR) plays a critical role at multiple stages of thymocyte differentiation, T-cell activation, and homeostasis [reviewed in (3;4)]. Syk and ZAP-70 function as critical mediators of pre-TCR and TCR signaling, with ZAP-70 having a predominant role in mature T cells (4;5). Once activated, ZAP-70 and Syk interact with and phosphorylate a number of substrates important for TCR signaling including the adaptor proteins the linker for activation of T cells (LAT) and SH2 domain-containing leukocyte protein of 76 kDa (SLP-76) (6;7). Once phosphorylated, these two adaptors serve as docking sites and organize a number of effector molecules into the correct spatiotemporal manner to allow the activation of multiple signaling pathways. Zap70 knockout mice display an arrest of T cell development at the DP stage, the second critical checkpoint important during αβ T cell development due to defective TCR-mediated selection and signaling at this stage (5;8). Although ZAP-70 has a critical role in T cell development and function, it also plays a role downstream of the BCR and in NK cells. Zap70 knockout mice display normal B cell development, mount normal antibody responses and also proliferate appropriately to various stimuli (9).
The wanna2 mice exhibit a similar phenotype to the wanna mice, which exhibit an ENU-induced mutation in Zap70. The phenotype of the wanna2 mouse provides in vivo confirmation that Cys 563 is important for ZAP70 function.
wanna2(F):5'- CCTTCTCCTATGGCCAGAAG -3'
wanna2(R):5'- TGTTGGGAAGCCAAGCACAG -3'
wanna2_seq(F):5'- GCCAGAAGCCCTACAAGGTAG -3'
wanna2_seq(R):5'- AAGAAGCCGGGTGTGACTCTG -3'
1. Adzhubei, I. A., Schmidt, S., Peshkin, L., Ramensky, V. E., Gerasimova, A., Bork, P., Kondrashov, A. S., and Sunyaev, S. R. (2010) A Method and Server for Predicting Damaging Missense Mutations. Nat Methods. 7, 248-249.
2. Au-Yeung, B. B., Deindl, S., Hsu, L. Y., Palacios, E. H., Levin, S. E., Kuriyan, J., and Weiss, A. (2009) The Structure, Regulation, and Function of ZAP-70. Immunol Rev. 228, 41-57.
3. Zamoyska, R., Basson, A., Filby, A., Legname, G., Lovatt, M., and Seddon, B. (2003) The Influence of the Src-Family Kinases, Lck and Fyn, on T Cell Differentiation, Survival and Activation. Immunol Rev. 191, 107-118.
4. Palacios, E. H., and Weiss, A. (2004) Function of the Src-Family Kinases, Lck and Fyn, in T-Cell Development and Activation. Oncogene. 23, 7990-8000.
5. Kadlecek, T. A., van Oers, N. S., Lefrancois, L., Olson, S., Finlay, D., Chu, D. H., Connolly, K., Killeen, N., and Weiss, A. (1998) Differential Requirements for ZAP-70 in TCR Signaling and T Cell Development. J Immunol. 161, 4688-4694.
6. Bubeck Wardenburg, J., Fu, C., Jackman, J. K., Flotow, H., Wilkinson, S. E., Williams, D. H., Johnson, R., Kong, G., Chan, A. C., and Findell, P. R. (1996) Phosphorylation of SLP-76 by the ZAP-70 Protein-Tyrosine Kinase is Required for T-Cell Receptor Function. J Biol Chem. 271, 19641-19644.
7. Zhang, W., Sloan-Lancaster, J., Kitchen, J., Trible, R. P., and Samelson, L. E. (1998) LAT: The ZAP-70 Tyrosine Kinase Substrate that Links T Cell Receptor to Cellular Activation. Cell. 92, 83-92.
8. Negishi, I., Motoyama, N., Nakayama, K., Nakayama, K., Senju, S., Hatakeyama, S., Zhang, Q., Chan, A. C., and Loh, D. Y. (1995) Essential Role for ZAP-70 in both Positive and Negative Selection of Thymocytes. Nature. 376, 435-438.
|Science Writers||Anne Murray|
|Authors||Jin Huk Choi, James Butler, Bruce Beutler|