Kinase Subfamily MAST

Kinase Classification: Group AGC: Family MAST: Subfamily MAST

MAST (Microtubule Associated Ser/Thr Kinases) are eukaryotic-wide kinases with roles in microtubule function, PTEN regulation and a variety of neuronal functions. They are targetted to substrate proteins by their PDZ domains.

Evolution

MAST kinases are found in most eukaryotes, though lost from most fungi and ciliates. They include single homologs in C. elegans (kin-4) and Drosophila (dop/CG6498) and four members (MAST1-4) in mammals.

Domain Structure

Animal MAST kinases have an N-terminal DUF1908 domain followed by a kinase domain with the AGC-specific tail extension, and a PDZ domain, with large unstructured regions in between. These accessory domains are found only in animal members. The DUF domain is of unknown function, and has only ever been found in MAST kinases. By analogy with other AGC kinases, it may regulate kinase activity. Two partial NMR structures (PDB:1V9V and PDB:2M9X) of DUF1908 from MAST1 and MAST3 show a 4-helix bundle arrangement, which is also seen in multiple alphafold predictions from the family.

Functions

Mammalian MAST1-3 can bind and phosphorylate the PTEN phosphatase via their PDZ domains which bind to the C-terminus of PTEN [1]. PTEN binds several other PDZ domains in a similar manner, so these proteins may compete for binding. The PTEN phosphorylation site is within the larger C-terminal region (350–403 in human), that includes a cluster of phosphorylation sites (380-385). Phosphorylation of this tail was reported to block PDZ binding and inhibit complex formation [2]. The rabies virus G protein mimics the PTEN C terminus and competes with PTEN for MAST2 binding. In cell culture, G protein caused nuclear exclusion of PTEN, likely as a result of blocked MAST2 binding [3]

Nematode kin-4 can also bind the C-terminus of PTEN via its PDZ domain in neurons and functions downstream of the daf-16 IGF1 receptor involved in longevity [4] (though C. elegans PTEN, daf-18, lacks a conserved C-terminal PDZ-interacting domain). It is also genetically implicated in thermotaxis in the AFD neuron, and genetically linked to mec-2 (a stomatin-like membrane-associated protein that binds microtubules) and diacyl glycerol kinase (dgk-1) [5].

Mutants in the Drosophila MAST, dop, have a defect in embryonic cellularization and have been implicated in both dynein and kinesin-based microtubule transport. dop is indicated to phosphorylate a conserved residue, S401, in Dlic, the Drosophila dynein light intermediate chain (homologous to human DYNC1LI1 and DYNC1LI2 (H. Sonnenberg thesis). dop also interacts genetically with dynein pathway genes and is required for phosphorylation of the dynein intermediate chain, sw [6].

High-throughput studies also link mammalian MASTs to cytoskelal motors including interactions between MAST2 and kinesin KIF23, the dynein-associated BICD2, tropomyosins 1-3, DYNNL1, and 3 tropomyosins (TPM1-3). MAST1 associates with two syntrophins (STNA1, STNB2). MAST3 interacts with kinesins (KIF1B, KIF1C, KIF13B), and MAST4 also interacts with BICD. All four interact with multiple 14-3-3 proteins.

Mouse Mast2 interacts with the dynein light chain, LC8 [7], and human MAST2 interacts with protocadherin KLC (CDHR2), via the MAST PDZ domain and the CDHR2 tail [8].

In Arabidopsis, MAST kinase IREH1 is named after its phenotype of incomplete root hair elongation [9]

MAST3 phosphorylates ARPP-16/19 in striatal neurons [10]. This is similar to the phosphorylation of ARPP-19 and ENSA by MASTL kinase, which turns them into inhibitors of the PP2A-B55 phosphatase complex, and promotes mitotic progression. PKA phosphorylates ARPP-16 in a way that blocks MAST3 phosphorylation and also phosphorylates MAST3, inhibiting it's kinase activity [11].

MASTs are associated with a number of human diseases, mostly neuronal. These include germline MAST1 mutations in mega-corpus-callosum syndrome (brain malformations) [12], MAST2 duplications with azoospermia [13], a MAST2 mutation with thrombophilia [14], and MAST3 and MAST4 mutations with epilepsy [15, 16].

MAST2 is involved in NFkB signaling, by binding TRAF6 and TRAF2 and blocking their activation of NFkB [17]. Binding involves the region 218-233, (upstream of the 4 helix bundle but within the DUF1608 domain). In macrophages and cell lines, MAST2 was implicated in NF-kB-mediated induction of IL-12 synthesis after LPS treatment [18]. MAST3 is also implicated in NF-kB signaling, by genetic variants linked to inflammatory bowel disease (IBD) and the finding that MAST3 knockdown decreased TLR4-dependent NF-kB activity [19]. Expression of a gene set responsive to altered MAST3 levels was elevated in IBD patients, and encoded genes involved in NF-kb signaling [20]. MAST3 was implicated in activation of NK-kB and Wnt signaling, as a target of MiR-125a-3p in fibroblast-like synovial cells in rheumatoid arthritis [21].

MAST1 and MAST2 interact with beta 2-syntrophin, a neuromuscular junction protein that binds dystophin via the PDZ domains of both proteins [22].

MAST4 is an estrogen-responsive gene whose expression in female multiple myeloma patients is increased, and reduces the incidence of MM bone disease, possibly by modulation of PTEN signaling [23].

MAST2 was first found as a testis-expressed protein that was microtubule-associated and could phosphorylate an unknown 75kd protein that was bound to it [24] Mouse Mast4 is required for spermatogenesis, and was shown to phosphorylate the transcription factor ERM (Ets-related molecule) on S367 (also a PKA site), which regulates expression of ERM target genes and self-renewal of spermatogonial stem cells.

MAST2 phosphorylates and inhibits Na+/H+ exchanger 3 (NHE3, SLC9A3); [25]. Binding is in the region C-terminal of the kinase domain and may involve the PDZ domain, as NHE3 is known to bind other PDZ domains [26], and the internal PDZ-binding motif is similar to that on PTEN. Inhibition was dependent on kinase activity.

In humans, MAST1 is almost exclusively expressed in brain, with some in testis, MAST2 widespread and highest in testis, cervix and muscle, MAST3 is widespread but high in brain, and MAST4 is expressed in most tissues, high in several endodermal-rich tissues (sourced from Entrez Gene, GTEx).

References

1. Valiente M, Andrés-Pons A, Gomar B, Torres J, Gil A, Tapparel C, Antonarakis SE, and Pulido R. Binding of PTEN to specific PDZ domains contributes to PTEN protein stability and phosphorylation by microtubule-associated serine/threonine kinases. J Biol Chem. 2005 Aug 12;280(32):28936-43. DOI:10.1074/jbc.M504761200 |
2. Vazquez F, Grossman SR, Takahashi Y, Rokas MV, Nakamura N, and Sellers WR. Phosphorylation of the PTEN tail acts as an inhibitory switch by preventing its recruitment into a protein complex. J Biol Chem. 2001 Dec 28;276(52):48627-30. DOI:10.1074/jbc.C100556200 |
3. Terrien E, Chaffotte A, Lafage M, Khan Z, Préhaud C, Cordier F, Simenel C, Delepierre M, Buc H, Lafon M, and Wolff N. Interference with the PTEN-MAST2 interaction by a viral protein leads to cellular relocalization of PTEN. Sci Signal. 2012 Aug 14;5(237):ra58. DOI:10.1126/scisignal.2002941 |
4. An SWA, Choi ES, Hwang W, Son HG, Yang JS, Seo K, Nam HJ, Nguyen NTH, Kim EJE, Suh BK, Kim Y, Nakano S, Ryu Y, Man Ha C, Mori I, Park SK, Yoo JY, Kim S, and Lee SV. KIN-4/MAST kinase promotes PTEN-mediated longevity of Caenorhabditis elegans via binding through a PDZ domain. Aging Cell. 2019 Jun;18(3):e12906. DOI:10.1111/acel.12906 |
5. Nakano S, Ikeda M, Tsukada Y, Fei X, Suzuki T, Niino Y, Ahluwalia R, Sano A, Kondo R, Ihara K, Miyawaki A, Hashimoto K, Higashiyama T, and Mori I. Presynaptic MAST kinase controls opposing postsynaptic responses to convey stimulus valence in Caenorhabditis elegans. Proc Natl Acad Sci U S A. 2020 Jan 21;117(3):1638-1647. DOI:10.1073/pnas.1909240117 |
6. Hain D, Langlands A, Sonnenberg HC, Bailey C, Bullock SL, and Müller HA. The Drosophila MAST kinase Drop out is required to initiate membrane compartmentalisation during cellularisation and regulates dynein-based transport. Development. 2014 May;141(10):2119-30. DOI:10.1242/dev.104711 |
7. Navarro-Lérida I, Martínez Moreno M, Roncal F, Gavilanes F, Albar JP, and Rodríguez-Crespo I. Proteomic identification of brain proteins that interact with dynein light chain LC8. Proteomics. 2004 Feb;4(2):339-46. DOI:10.1002/pmic.200300528 |
8. Okazaki N, Takahashi N, Kojima S, Masuho Y, and Koga H. Protocadherin LKC, a new candidate for a tumor suppressor of colon and liver cancers, its association with contact inhibition of cell proliferation. Carcinogenesis. 2002 Jul;23(7):1139-48. DOI:10.1093/carcin/23.7.1139 |
9. Chudinova EM, Karpov PA, Fokin AI, Yemets AI, Lytvyn DI, Nadezhdina ES, and Blume YB. MAST-like protein kinase IREH1 from Arabidopsis thaliana co-localizes with the centrosome when expressed in animal cells. Planta. 2017 Nov;246(5):959-969. DOI:10.1007/s00425-017-2742-4 |
10. Andrade EC, Musante V, Horiuchi A, Matsuzaki H, Brody AH, Wu T, Greengard P, Taylor JR, and Nairn AC. ARPP-16 Is a Striatal-Enriched Inhibitor of Protein Phosphatase 2A Regulated by Microtubule-Associated Serine/Threonine Kinase 3 (Mast 3 Kinase). J Neurosci. 2017 Mar 8;37(10):2709-2722. DOI:10.1523/JNEUROSCI.4559-15.2017 |
11. Musante V, Li L, Kanyo J, Lam TT, Colangelo CM, Cheng SK, Brody AH, Greengard P, Le Novère N, and Nairn AC. Reciprocal regulation of ARPP-16 by PKA and MAST3 kinases provides a cAMP-regulated switch in protein phosphatase 2A inhibition. Elife. 2017 Jun 14;6. DOI:10.7554/eLife.24998 |
12. Tripathy R, Leca I, van Dijk T, Weiss J, van Bon BW, Sergaki MC, Gstrein T, Breuss M, Tian G, Bahi-Buisson N, Paciorkowski AR, Pagnamenta AT, Wenninger-Weinzierl A, Martinez-Reza MF, Landler L, Lise S, Taylor JC, Terrone G, Vitiello G, Del Giudice E, Brunetti-Pierri N, D'Amico A, Reymond A, Voisin N, Bernstein JA, Farrelly E, Kini U, Leonard TA, Valence S, Burglen L, Armstrong L, Hiatt SM, Cooper GM, Aldinger KA, Dobyns WB, Mirzaa G, Pierson TM, Baas F, Chelly J, Cowan NJ, and Keays DA. Mutations in MAST1 Cause Mega-Corpus-Callosum Syndrome with Cerebellar Hypoplasia and Cortical Malformations. Neuron. 2018 Dec 19;100(6):1354-1368.e5. DOI:10.1016/j.neuron.2018.10.044 |
13. pmid= 26203179
14. Morange PE, Peiretti F, Gourhant L, Proust C, Soukarieh O, Pulcrano-Nicolas AS, Saripella GV, Stefanucci L, Lacroix R, Ibrahim-Kosta M, Lemarié CA, Frontini M, Alessi MC, Trégouët DA, and Couturaud F. A rare coding mutation in the MAST2 gene causes venous thrombosis in a French family with unexplained thrombophilia: The Breizh MAST2 Arg89Gln variant. PLoS Genet. 2021 Jan;17(1):e1009284. DOI:10.1371/journal.pgen.1009284 |
15. Landoulsi Z, Laatar F, Noé E, Mrabet S, Ben Djebara M, Achaz G, Nava C, Baulac S, Kacem I, Gargouri-Berrechid A, Gouider R, and Leguern E. Clinical and genetic study of Tunisian families with genetic generalized epilepsy: contribution of CACNA1H and MAST4 genes. Neurogenetics. 2018 Aug;19(3):165-178. DOI:10.1007/s10048-018-0550-z |
16. Spinelli E, Christensen KR, Bryant E, Schneider A, Rakotomamonjy J, Muir AM, Giannelli J, Littlejohn RO, Roeder ER, Schmidt B, Wilson WG, Marco EJ, Iwama K, Kumada S, Pisano T, Barba C, Vetro A, Brilstra EH, van Jaarsveld RH, Matsumoto N, Goldberg-Stern H, Carney PW, Andrews PI, El Achkar CM, Berkovic S, Rodan LH, Undiagnosed Diseases Network (UDN), McWalter K, Guerrini R, Scheffer IE, Mefford HC, Mandelstam S, Laux L, Millichap JJ, Guemez-Gamboa A, Nairn AC, and Carvill GL. Pathogenic MAST3 Variants in the STK Domain Are Associated with Epilepsy. Ann Neurol. 2021 Aug;90(2):274-284. DOI:10.1002/ana.26147 |
17. Xiong H, Li H, Chen Y, Zhao J, and Unkeless JC. Interaction of TRAF6 with MAST205 regulates NF-kappaB activation and MAST205 stability. J Biol Chem. 2004 Oct 15;279(42):43675-83. DOI:10.1074/jbc.M404328200 |
18. Zhou H, Xiong H, Li H, Plevy SE, Walden PD, Sassaroli M, Prestwich GD, and Unkeless JC. Microtubule-associated serine/threonine kinase-205 kDa and Fc gamma receptor control IL-12 p40 synthesis and NF-kappa B activation. J Immunol. 2004 Feb 15;172(4):2559-68. DOI:10.4049/jimmunol.172.4.2559 |
19. Labbé C, Goyette P, Lefebvre C, Stevens C, Green T, Tello-Ruiz MK, Cao Z, Landry AL, Stempak J, Annese V, Latiano A, Brant SR, Duerr RH, Taylor KD, Cho JH, Steinhart AH, Daly MJ, Silverberg MS, Xavier RJ, and Rioux JD. MAST3: a novel IBD risk factor that modulates TLR4 signaling. Genes Immun. 2008 Oct;9(7):602-12. DOI:10.1038/gene.2008.57 |
20. Labbé C, Boucher G, Foisy S, Alikashani A, Nkwimi H, David G, Beaudoin M, Goyette P, Charron G, Xavier RJ, and Rioux JD. Genome-wide expression profiling implicates a MAST3-regulated gene set in colonic mucosal inflammation of ulcerative colitis patients. Inflamm Bowel Dis. 2012 Jun;18(6):1072-80. DOI:10.1002/ibd.21887 |
21. Wang Y, Yin Z, Zhang N, Song H, Zhang Q, Hao X, and Wang Z. MiR-125a-3p inhibits cell proliferation and inflammation responses in fibroblast-like synovial cells in rheumatoid arthritis by mediating the Wnt/β-catenin and NF-κB pathways via targeting MAST3. J Musculoskelet Neuronal Interact. 2021 Dec 1;21(4):560-567.
22. Lumeng C, Phelps S, Crawford GE, Walden PD, Barald K, and Chamberlain JS. Interactions between beta 2-syntrophin and a family of microtubule-associated serine/threonine kinases. Nat Neurosci. 1999 Jul;2(7):611-7. DOI:10.1038/10165 |
23. Cui Y, Wang F, Zhang D, Huang J, Yang Y, Xu J, Gao Y, Ding H, Qu Y, Zhang W, Liu W, Pan L, Zhang L, Liu Z, Niu T, Liu T, and Zheng Y. Estrogen-Responsive Gene MAST4 Regulates Myeloma Bone Disease. J Bone Miner Res. 2022 Apr;37(4):711-723. DOI:10.1002/jbmr.4507 |
24. Walden PD and Cowan NJ. A novel 205-kilodalton testis-specific serine/threonine protein kinase associated with microtubules of the spermatid manchette. Mol Cell Biol. 1993 Dec;13(12):7625-35. DOI:10.1128/mcb.13.12.7625-7635.1993 |
25. Wang D, Lee HJ, Cooper DS, Cebotaro L, Walden PD, Choi I, and Yun CC. Coexpression of MAST205 inhibits the activity of Na+/H+ exchanger NHE3. Am J Physiol Renal Physiol. 2006 Feb;290(2):F428-37. DOI:10.1152/ajprenal.00161.2005 |
26. Cha B, Yang J, Singh V, Zachos NC, Sarker RI, Chen TE, Chakraborty M, Tse CM, and Donowitz M. PDZ domain-dependent regulation of NHE3 protein by both internal Class II and C-terminal Class I PDZ-binding motifs. J Biol Chem. 2017 May 19;292(20):8279-8290. DOI:10.1074/jbc.M116.774489 |