Abstract
To gain a better understanding of how Crk II regulates the function of the Abl tyrosine kinase, we explored the function of the C-terminal linker and SH3 domain, a region of Crk II that is still poorly understood. Molecular modeling, tryptophan fluorescence, and covariation sequence alignment indicate that the Crk-SH3-C has a unique binding groove and RT loop not observed in typical SH3 domains. Based on these models, we made a series of mutations in the linker and in residues predicted to destabilize the putative binding pocket and RT loop. In Abl transactivation assays, Y222F and P225A mutations in the linker resulted in strong transactivation of Abl by Crk II. However, mutations predicted to be at the surface of the Crk SH3-C were not activators of Abl. Interestingly, combinations of activating mutations of Crk II with mutations in the highly conserved PNAY sequence in the SH3-C inactivated the activating mutations, suggesting that the SH3-C is necessary for activation. Our data provide insight into the role of highly conserved residues in the Crk-SH3-C, suggesting a mechanism for how the linker and the Crk-SH3-C function in the transactivation of the Abl tyrosine kinase.
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References
Ago T, Nunoi H, Ito T and Sumimoto H . (1999). J. Biol. Chem., 274, 33644–33653.
Barber DL, Beattie BK, Mason JM, Nguyen MH, Yoakim M, Neel BG, D'Andrea AD and Frank DA . (2001). Blood, 97, 2230–2237.
Barnett P, Bottger G, Klein AT, Tabak HF and Distel B . (2000). EMBO J., 19, 6382–6391.
Brasher BB and Van Etten RA . (2000). J. Biol. Chem., 275, 35631–35637.
Evans EK, Lu W, Strum SL, Mayer BJ and Kornbluth S . (1997). EMBO J., 16, 230–241.
Feller SM . (2001). Oncogene, 20, 6348–6371.
Feller SM, Knudsen B and Hanafusa H . (1994). EMBO J., 13, 2341–2351.
Feller SM, Knudsen B, Wong TW and Hanafusa H . (1995). Methods Enzymol., 255, 369–378.
Galletta BJ, Niu XP, Erickson MR and Abmayr SM . (1999). Gene, 228, 243–252.
Hantschel O and Superti-Furga G . (2004). Nat. Rev. Mol. Cell Biol., 5, 33–44.
Harkiolaki M, Lewitzky M, Gilbert RJ, Jones EY, Bourette RP, Mouchiroud G, Sondermann H, Moarefi I and Feller SM . (2003). EMBO J., 22, 2571–2582.
Hemmeryckx B, Reichert A, Watanabe M, Kaartinen V, de Jong R, Pattengale PK, Groffen J and Heisterkamp N . (2002). Oncogene, 21, 3225–3231.
Kami K, Takeya R, Sumimoto H and Kohda D . (2002). EMBO J., 21, 4268–4276.
Kang H, Freund C, Duke-Cohan JS, Musfelleracchio A, Wagner G and Rudd CE . (2000). EMBO J., 19, 2889–2899.
Kay BK, Williamson MP and Sudol M . (2000). FASEB J., 14, 231–241.
Kelley LA, MacCallum RM and Sternberg MJ . (2000). J. Mol. Biol., 299, 499–520.
Kizaka-Kondoh S, Matsuda M and Okayama H . (1996). Proc. Natl. Acad. Sci. USA, 93, 12177–12182.
Knudsen BS, Feller SM and Hanafusa H . (1994). J. Biol. Chem., 269, 32781–32787.
Kurakin A and Bredesen D . (2002). J. Biomol. Struct. Dyn., 19, 1015–1029.
Kurakin AV, Wu S and Bredesen DE . (2003). J. Biol. Chem., 278, 34102–34109. Epub 2003 June 26.
Larson SM and Davidson AR . (2000). Protein Sci., 9, 2170–2180.
Larson SM, Di Nardo AA and Davidson AR . (2000). J. Mol. Biol., 303, 433–446.
Lennon G, Auffray C, Polymeropoulos M and Soares MB . (1996). Genomics, 33, 151–152.
Lewitzky M, Harkiolaki M, Domart MC, Jones EY and Feller SM . (2004). J. Biol. Chem., 279, 28724–28732 Epub 2004 April 20..
Lewitzky M, Kardinal C, Gehring NH, Schmidt EK, Konkol B, Eulitz M, Birchmeier W, Schaeper U and Feller SM . (2001). Oncogene, 20, 1052–1062.
Lowenstein EJ, Daly RJ, Batzer AG, Li W, Margolis B, Lammers R, Ullrich A, Skolnik EY, Bar-Sagi D and Schlessinger J . (1992). Cell, 70, 431–442.
Macias MJ, Wiesner S and Sudol M . (2002). FEBS Lett., 513, 30–37.
Matsuda M, Tanaka S, Nagata S, Kojima A, Kurata T and Shibuya M . (1992). Mol. Cell. Biol., 12, 3482–3489.
Mayer BJ, Hamaguchi M and Hanafusa H . (1988). Nature, 332, 272–275.
Mongiovi AM, Romano PR, Panni S, Mendoza M, Wong WT, Musacchio A, Cesareni G and Di Fiore PP . (1999). EMBO J., 18, 5300–5309.
Nagar B, Hantschel O, Young MA, Scheffzek K, Veach D, Bornmann W, Clarkson B, Superti-Furga G and Kuriyan J . (2003). Cell, 112, 859–871.
Ogawa S, Toyoshima H, Kozutsumi H, Hagiwara K, Sakai R, Tanaka T, Hirano N, Mano H, Yazaki Y and Hirai H . (1994). Oncogene, 9, 1669–1678.
Pires JR, Hong X, Brockmann C, Volkmer-Engert R, Schneider-Mergener J, Oschkinat H and Erdmann R . (2003). J. Mol. Biol., 326, 1427–1435.
Reddien PW and Horvitz HR . (2000). Nat. Cell. Biol., 2, 131–136.
Reichman CT, Mayer BJ, Keshav S and Hanafusa H . (1992). Cell Growth Differ., 3, 451–460.
Ren R, Ye ZS and Baltimore D . (1994). Genes Dev., 8, 783–795.
Rosen MK, Yamazaki T, Gish GD, Kay CM, Pawson T and Kay LE . (1995). Nature, 374, 477–479.
Rozakis-Adcock M, McGlade J, Mbamalu G, Pelicci G, Daly R, Li W, Batzer A, Thomas S, Brugge J, Pelicci PC, Schlessinger J and Pawson T . (1992). Nature, 360, 689–692.
Sattler M and Salgia R . (1998). Leukemia, 12, 637–644.
Shi J, Blundell TL and Mizuguchi K . (2001). J. Mol. Biol., 310, 243–257.
Shishido T, Akagi T, Chalmers A, Maeda M, Terada T, Georgescu MM and Hanafusa H . (2001). Genes Cells, 6, 431–440.
Sicheri F, Moarefi I and Kuriyan J . (1997). Nature, 385, 602–609.
Sippl MJ . (1993). Proteins, 17, 355–362.
Smith JJ, Richardson DA, Kopf J, Yoshida M, Hollingsworth RE and Kornbluth S . (2002). Mol. Cell. Biol., 22, 1412–1423.
Smith KM and Van Etten RA . (2001). J. Biol. Chem., 276, 24372–24379.
Songyang Z, Shoelson SE, Chaudhuri M, Gish G, Pawson T, Haser WG, King F, Roberts T, Ratnofsky S, Lechleider RJ, Neel B, Birge RB, Fajardo JE, Chou MM, Hanafusa H, Schaffhausen B and Cantley LC . (1993). Cell, 72, 767–778.
Tanis KQ, Veach D, Duewel HS, Bornmann WG and Koleske AJ . (2003). Mol. Cell. Biol., 23, 3884–3896.
ten Hoeve J, Morris C, Heisterkamp N and Groffen J . (1993). Oncogene, 8, 2469–2474.
Van Etten RA, Jackson P and Baltimore D . (1989). Cell, 58, 669–678.
Wang JY . (2004). Nat. Cell Biol., 6, 3–7.
Wang B, Mysliwiec T, Feller SM, Knudsen B, Hanafusa H and Kruh GD . (1996). Oncogene, 13, 1379–1385.
Wang W and Malcolm BA . (1999). BioTechniques, 26, 680–682.
Wu X, Knudsen B, Feller SM, Zheng J, Sali A, Cowburn D, Hanafusa H and Kuriyan J . (1995). Structure, 3, 215–226.
Zvara A, Fajardo JE, Escalante M, Cotton G, Muir T, Kirsch KH and Birge RB . (2001). Oncogene, 20, 951–961.
Acknowledgements
We thank Drs Cecile Bougeret, Brain Kay, Tom Muir, Stephan Feller, Sally Kornbluth, and Allan Davidson for insightful discussions and sharing unpublished data. This work was supported by a Public Health Service Award (NIH-GMS55760) to RBB and an NJCCC fellowship to Sukhwinder Singh.
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Reichman, C., Singh, K., Liu, Y. et al. Transactivation of Abl by the Crk II adapter protein requires a PNAY sequence in the Crk C-terminal SH3 domain. Oncogene 24, 8187–8199 (2005). https://doi.org/10.1038/sj.onc.1208988
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DOI: https://doi.org/10.1038/sj.onc.1208988
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