Regulation of Src tumor activity by its N-terminal intrinsically disordered region.
Journal
Oncogene
ISSN: 1476-5594
Titre abrégé: Oncogene
Pays: England
ID NLM: 8711562
Informations de publication
Date de publication:
02 2022
02 2022
Historique:
received:
22
04
2021
accepted:
21
10
2021
revised:
14
10
2021
pubmed:
10
1
2022
medline:
24
2
2022
entrez:
9
1
2022
Statut:
ppublish
Résumé
The membrane-anchored Src tyrosine kinase is involved in numerous pathways and its deregulation is involved in human cancer. Our knowledge on Src regulation relies on crystallography, which revealed intramolecular interactions to control active Src conformations. However, Src contains a N-terminal intrinsically disordered unique domain (UD) whose function remains unclear. Using NMR, we reported that UD forms an intramolecular fuzzy complex involving a conserved region with lipid-binding capacity named Unique Lipid-Binding Region (ULBR), which could modulate Src membrane anchoring. Here we show that the ULBR is essential for Src's oncogenic capacity. ULBR inactive mutations inhibited Src transforming activity in NIH3T3 cells and in human colon cancer cells. It also reduced Src-induced tumor development in nude mice. An intact ULBR was required for MAPK signaling without affecting Src kinase activity nor sub-cellular localization. Phospho-proteomic analyses revealed that, while not impacting on the global tyrosine phospho-proteome in colon cancer cells, this region modulates phosphorylation of specific membrane-localized tyrosine kinases needed for Src oncogenic signaling, including EPHA2 and Fyn. Collectively, this study reveals an important role of this intrinsically disordered region in malignant cell transformation and suggests a novel layer of Src regulation by this unique region via membrane substrate phosphorylation.
Identifiants
pubmed: 34999732
doi: 10.1038/s41388-021-02092-x
pii: 10.1038/s41388-021-02092-x
pmc: PMC8837538
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
960-970Commentaires et corrections
Type : ErratumIn
Informations de copyright
© 2021. The Author(s).
Références
Yeatman TJ. A renaissance for SRC. Nat Rev Cancer. 2004;4:470–80.
doi: 10.1038/nrc1366
Summy JM, Gallick GE. Src family kinases in tumor progression and metastasis. Cancer Metastasis Rev. 2003;22:337–58.
doi: 10.1023/A:1023772912750
Sirvent A, Mevizou R, Naim D, Lafitte M, Roche S. Src family tyrosine kinases in intestinal homeostasis, regeneration and tumorigenesis. Cancers. 2020;12:2014.
doi: 10.3390/cancers12082014
Boggon TJ, Eck MJ. Structure and regulation of Src family kinases. Oncogene. 2004;23:7918–27.
doi: 10.1038/sj.onc.1208081
Arbesú M, Iruela G, Fuentes H, Teixeira JMC, Pons M. Intramolecular fuzzy interactions involving intrinsically disordered domains. Front Mol Biosci. 2018;5:39.
doi: 10.3389/fmolb.2018.00039
Amata I, Maffei M, Pons M. Phosphorylation of unique domains of Src family kinases. Front Genet. 2014;5. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4075076/ .
Cross FR, Garber EA, Pellman D, Hanafusa H. A short sequence in the p60src N terminus is required for p60src myristylation and membrane association and for cell transformation. Mol Cell Biol. 1984;4:1834–42.
pubmed: 6092942
pmcid: 368993
Wright PE, Dyson HJ. Intrinsically disordered proteins in cellular signaling and regulation. Nat Rev Mol Cell Biol. 2015;16:18–29.
doi: 10.1038/nrm3920
Fuxreiter M. Fuzziness: linking regulation to protein dynamics. Mol BioSyst. 2011;8:168–77. 1
doi: 10.1039/C1MB05234A
Pérez Y, Maffei M, Igea A, Amata I, Gairí M, Nebreda AR, et al. Lipid binding by the Unique and SH3 domains of c-Src suggests a new regulatory mechanism. Sci Rep. 2013;3:1295.
doi: 10.1038/srep01295
Maffei M, Arbesú M, Le Roux A-L, Amata I, Roche S, Pons M. The SH3 domain acts as a scaffold for the N-terminal intrinsically disordered regions of c-Src. Structure. 2015;23:893–902. 5
doi: 10.1016/j.str.2015.03.009
Spassov DS, Ruiz-Saenz A, Piple A, Moasser MM. A dimerization function in the intrinsically disordered N-terminal region of Src. Cell Rep. 2018;25:449–463.e4. 9
doi: 10.1016/j.celrep.2018.09.035
Le Roux A-L, Mohammad I-L, Mateos B, Arbesú M, Gairí M, Khan FA, et al. A myristoyl-binding site in the SH3 domain modulates c-Src membrane anchoring. iScience. 2019;12:194–203. 22
doi: 10.1016/j.isci.2019.01.010
Arbesú M, Maffei M, Cordeiro TN, Teixeira JMC, Pérez Y, Bernadó P, et al. The unique domain forms a fuzzy intramolecular complex in Src family kinases. Structure. 2017;25:630–640.e4. 4
doi: 10.1016/j.str.2017.02.011
Teixeira JMC, Fuentes H, Bielskutė S, Gairi M, Żerko S, Koźmiński W, et al. The two isoforms of lyn display different intramolecular fuzzy complexes with the SH3 domain. Molecules. 2018;23:2731.
Pérez Y, Gairí M, Pons M, Bernadó P. Structural characterization of the natively unfolded N-terminal domain of human c-Src kinase: insights into the role of phosphorylation of the unique domain. J Mol Biol. 2009;391:136–48. 7
doi: 10.1016/j.jmb.2009.06.018
Amata I, Maffei M, Igea A, Gay M, Vilaseca M, Nebreda AR, et al. Multi-phosphorylation of the intrinsically disordered unique domain of c-Src studied by in-cell and real-time NMR spectroscopy. ChemBioChem. 2013;14:1820–7.
doi: 10.1002/cbic.201300139
Yokouchi M, Kondo T, Sanjay A, Houghton A, Yoshimura A, Komiya S, et al. Src-catalyzed phosphorylation of c-Cbl leads to the interdependent ubiquitination of both proteins. J Biol Chem. 2001;276:35185–93. 14
doi: 10.1074/jbc.M102219200
Manes G, Bello P, Roche S. Slap negatively regulates Src mitogenic function but does not revert Src-induced cell morphology changes. Mol Cell Biol. 2000;20:3396–406.
doi: 10.1128/MCB.20.10.3396-3406.2000
Sirvent A, Bénistant C, Pannequin J, Veracini L, Simon V, Bourgaux J-F, et al. Src family tyrosine kinases-driven colon cancer cell invasion is induced by Csk membrane delocalization. Oncogene. 2010;29:1303–15. 4
doi: 10.1038/onc.2009.450
Naudin C, Sirvent A, Leroy C, Larive R, Simon V, Pannequin J, et al. SLAP displays tumour suppressor functions in colorectal cancer via destabilization of the SRC substrate EPHA2. Nat Commun. 2014;5:3159.
doi: 10.1038/ncomms4159
Leroy C, Fialin C, Sirvent A, Simon V, Urbach S, Poncet J, et al. Quantitative phosphoproteomics reveals a cluster of tyrosine kinases that mediates SRC invasive activity in advanced colon carcinoma cells. Cancer Res. 2009;69:2279–86. 15
doi: 10.1158/0008-5472.CAN-08-2354
Sirvent A, Vigy O, Orsetti B, Urbach S, Roche S. Analysis of SRC oncogenic signaling in colorectal cancer by stable isotope labeling with heavy amino acids in mouse xenografts. Mol Cell Proteom. 2012;11:1937–50.
doi: 10.1074/mcp.M112.018168
Lecointre C, Simon V, Kerneur C, Allemand F, Fournet A, Montarras I, et al. Dimerization of the pragmin pseudo-kinase regulates protein tyrosine phosphorylation. Structure. 2018;26:545–554.e4. 3
doi: 10.1016/j.str.2018.01.017
Sirvent A, Benistant C, Roche S. Oncogenic signaling by tyrosine kinases of the SRC family in advanced colorectal cancer. Am J Cancer Res. 2012;2:357–71.
pubmed: 22860228
pmcid: 3410585
Turkson J, Bowman T, Adnane J, Zhang Y, Djeu JY, Sekharam M, et al. Requirement for Ras/Rac1-mediated p38 and c-Jun N-terminal kinase signaling in Stat3 transcriptional activity induced by the Src oncoprotein. Mol Cell Biol. 1999;19:7519–28.
doi: 10.1128/MCB.19.11.7519
Bowman T, Broome MA, Sinibaldi D, Wharton W, Pledger WJ, Sedivy JM, et al. Stat3-mediated Myc expression is required for Src transformation and PDGF-induced mitogenesis. Proc Natl Acad Sci USA. 2001;98:7319–24. 19
doi: 10.1073/pnas.131568898
Le Roux A-L, Busquets MA, Sagués F, Pons M. Kinetics characterization of c-Src binding to lipid membranes: Switching from labile to persistent binding. Colloids Surf B Biointerfaces. 2016;138:17–25. 1
doi: 10.1016/j.colsurfb.2015.11.013
Veracini L, Franco M, Boureux A, Simon V, Roche S, Benistant C. Two distinct pools of Src family tyrosine kinases regulate PDGF-induced DNA synthesis and actin dorsal ruffles. J Cell Sci. 2006;119:2921–34. 15
doi: 10.1242/jcs.03015
Le Roux A-L, Castro B, Garbacik ET, Garcia Parajo MF, Pons M. Single molecule fluorescence reveals dimerization of myristoylated Src N-terminal region on supported lipid bilayers. ChemistrySelect. 2016;1:642–7. 1
doi: 10.1002/slct.201600117
Jeitany M, Leroy C, Tosti P, Lafitte M, Le Guet J, Simon V, et al. Inhibition of DDR1-BCR signalling by nilotinib as a new therapeutic strategy for metastatic colorectal cancer. EMBO Mol Med. 2018;10:e7918.
Kashiwagi K, Ito S, Maeda S, Kato G. A Ser75-to-Asp phospho-mimicking mutation in Src accelerates ageing-related loss of retinal ganglion cells in mice. Sci Rep. 2017;7:16779. 1
doi: 10.1038/s41598-017-16872-7
Kato G. Nonphosphorylatable Src Ser75 mutation increases ethanol preference and consumption in mice. eNeuro. 2019;6:e0418–18.2019 1–1.
Kerjouan A, Boyault C, Oddou C, Hiriart-Bryant E, Grichine A, Kraut A, et al. Control of SRC molecular dynamics encodes distinct cytoskeletal responses by specifying signaling pathway usage. J Cell Sci. 2021;134:jcs254599.
doi: 10.1242/jcs.254599
Kim PW, Sun Z-YJ, Blacklow SC, Wagner G, Eck MJ. A zinc clasp structure tethers Lck to T cell coreceptors CD4 and CD8. Science. 2003;301:1725–8. 19
doi: 10.1126/science.1085643
Yu XM, Askalan R, Keil GJ, Salter MW. NMDA channel regulation by channel-associated protein tyrosine kinase Src. Science. 1997;275:674–8. 31
doi: 10.1126/science.275.5300.674
Nagar B, Hantschel O, Young MA, Scheffzek K, Veach D, Bornmann W, et al. Structural basis for the autoinhibition of c-Abl tyrosine kinase. Cell. 2003;112:859–71. 21
doi: 10.1016/S0092-8674(03)00194-6
Eide CA, Zabriskie MS, Savage Stevens SL, Antelope O, Vellore NA, Than H, et al. Combining the allosteric inhibitor asciminib with ponatinib suppresses emergence of and restores efficacy against highly resistant BCR-ABL1 mutants. Cancer Cell. 2019;36:431–443.e5. 14
doi: 10.1016/j.ccell.2019.08.004
Roche S, Fumagalli S, Courtneidge SA. Requirement for Src family protein tyrosine kinases in G2 for fibroblast cell division. Science. 1995;269:1567–9. 15
doi: 10.1126/science.7545311
Katoh K, Standley DM. MAFFT multiple sequence alignment software version 7: improvements in performance and usability. Mol Biol Evol. 2013;30:772–80.
doi: 10.1093/molbev/mst010
Guindon S, Gascuel O. A simple, fast, and accurate algorithm to estimate large phylogenies by maximum likelihood. Syst Biol. 2003;52:696–704.
doi: 10.1080/10635150390235520
Yang Z. PAML 4: phylogenetic analysis by maximum likelihood. Mol Biol Evol. 2007;24:1586–91.
doi: 10.1093/molbev/msm088