|Coordinate||36,779,173 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 (+)|
FUNCTION: This gene encodes a member of the protein tyrosine kinase family. The encoded protein is essential for development of T lymphocytes and thymocytes, and functions in the initial step of T lymphocyte receptor-mediated signal transduction. A mutation in this gene causes chronic autoimmune arthritis, similar to rheumatoid arthritis in humans. Mice lacking this gene are deficient in alpha-beta T lymphocytes in the thymus. In humans, mutations in this gene cause selective T-cell defect, a severe combined immunodeficiency disease characterized by a selective absence of CD8-positive T lymphocytes. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jan 2014]
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. [provided by MGI curators]
|Amino Acid Change||Tyrosine changed to Histidine|
|Institutional Source||Beutler Lab|
|Gene Model||predicted gene model for protein(s): [ENSMUSP00000027291]|
AA Change: Y314H
|Predicted Effect||probably damaging
PolyPhen 2 Score 0.998 (Sensitivity: 0.27; Specificity: 0.99)
|Meta Mutation Damage Score||0.0775|
|Is this an essential gene?||Probably nonessential (E-score: 0.233)|
|Candidate Explorer Status||CE: excellent candidate; human score: 5.5; ML prob: 0.63|
Linkage Analysis Data
|Alleles Listed at MGI|
|Mode of Inheritance||Autosomal Recessive|
|Last Updated||2019-09-04 9:42 PM by Diantha La Vine|
|Record Created||2016-10-21 7:35 AM|
The mesa_verde phenotype was identified by position-based superpedigree analysis of pedigrees R3689 and R3690. Homozygous mice in these pedigrees exhibited increased frequencies of CD44+ T cells (Figure 1) and CD44+ CD8 T cells (Figure 2), increased CD44 expression on T cells (Figure 3) and CD8+ T cells (Figure 4), and an increased frequency of central memory CD8+ T cells in CD8+ T cells (Figure 5). The response to the T-dependent antigen recombinant Semliki Forest virus (rSFV)-encoded β-galactosidase (rSFV-β-gal) was diminished (Figure 6).
|Nature of Mutation|
Whole exome HiSeq sequencing of the G1 grandsire identified 57 mutations in R3689 and 56 mutations in R3690. All of the above anomalies were linked by continuous variable mapping and position-based superpedigree analysis to a mutation in Zap70: an T to C transition at base pair 36,779,173 (v38) on chromosome 1, or base pair 17,376 in the GenBank genomic region NC_000067 encoding Zap70. The strongest association was found with a recessive model of linkage to the normalized antibody response to rSFV-β-gal, wherein three variant homozygotes departed phenotypically from 18 homozygous reference mice and 29 heterozygous mice with a P value of 5.432 x 10-10 (Figure 7). A substantial semidominant effect was observed in some of the assays but the mutation is preponderantly recessive, and in no assay was a purely dominant effect observed.
The mutation corresponds to residue 1,102 in the mRNA sequence NM_001289766 within exon 2 of 13 total exons.
The mutated nucleotide is indicated in red. The mutation results in a tyrosine (Y) to histidine (H) substitution at position 314 (Y314H) in the ZAP70 protein, and is strongly predicted by PolyPhen-2 to be damaging (score = 0.998) (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 mesa_verde mutation results in a tyrosine (Y) to histidine (H) substitution at position 314 (Y314H); residue 314 is within the IBD domain (Figure 8).
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 phenotype of the mesa_verde mice is similar to loss-of-function alleles of Zap70.
1) 94°C 2:00
The following sequence of 427 nucleotides is amplified (chromosome 1, + strand):
1 tgggtacatc atggcttatg gccgtgtgtc tggtgtgtct gagcttcccg tgtcctttac
Primer binding sites are underlined and the sequencing primers are highlighted; the mutated nucleotide is shown in red.
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|
|Illustrators||Peter Jurek, Katherine Timer|
|Authors||Ming Zeng, Xue Zhong, Jin Huk Choi, James Butler, and Bruce Beutler|