Skip to main content

Thank you for visiting You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

Src-mediated phosphorylation regulates subcellular distribution and activity of human inducible nitric oxide synthase


Inducible nitric oxide synthase (iNOS) expression is regulated at both the transcriptional and post-transcriptional level in epithelial cells. The aim of this study was to characterize the effects of tyrosine phosphorylation on iNOS activity. In a human intestinal epithelial cell line stimulated with cytokines, tyrosine phosphorylation of human iNOS protein was observed after 30 min exposure to pervanadate (PV), an inhibitor of protein tyrosine phosphatases. 4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine, a specific inhibitor of Src tyrosine kinases, abolished the PV-induced iNOS tyrosine phosphorylation. Cotransfection of Src with iNOS cDNA in human embryonic kidney (HEK) 293 cells resulted in a threefold (P<0.001) increase of iNOS protein levels and tyrosine phosphorylation of iNOS. In the presence of Src, 76% of wild-type (wt) iNOS was redistributed to detergent-insoluble domains and iNOS activity was decreased by 28% (P<0.05) despite increased iNOS protein levels. Analysis of iNOS tyrosine mutants revealed decreased Src-induced effects in Y151F iNOS mutant. Using a GST-fusion protein containing a domain encompassing Y151, we show that Y151 is a direct substrate for active Src in vitro. These findings indicate a role for iNOS tyrosine phosphorylation in the regulation of iNOS activity and the implication of Src tyrosine kinases in this pathway.

Your institute does not have access to this article

Access options

Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
Figure 7
Figure 8


  • Abram CL, Courtneidge SA . (2000). Exp Cell Res 254: 1–13.

  • Albakri QA, Stuehr DJ . (1996). J Biol Chem 271: 5414–5421.

  • Ambs S, Merriam WG, Bennett WP, Felley-Bosco E, Ogunfusika MO, Oser SM et al. (1998). Cancer Res 58: 334–341.

  • Andre M, Felley-Bosco E . (2003). FEBS Lett 546: 223–227.

  • Andre M, Latado H, Felley-Bosco E . (2005). Biochem Pharmacol 69: 595–602.

  • Baek KJ, Thiel BA, Lucas S, Stuehr DJ . (1993). J Biol Chem 268: 21120–21129.

  • Chiu T, Wu SS, Santiskulvong C, Tangkijvanich P, Yee Jr HF, Rozengurt E . (2002). Am J Physiol Cell Physiol 282: C434–C450.

  • Cobbs CS, Brenman JE, Aldape KD, Bredt DS, Israel MA . (1995). Cancer Res 55: 727–730.

  • Espey MG, Miranda KM, Pluta RM, Wink DA . (2000). J Biol Chem 275: 11341–11347.

  • Felley-Bosco E, Ambs S, Lowenstein CJ, Keefer LK, Harris CC . (1994). Am J Respir Cell Mol Biol 11: 159–164.

  • Felley-Bosco E, Bender FC, Courjault-Gautier F, Bron C, Quest AFG . (2000). Proc Natl Acad Sci USA 97: 14334–14339.

  • Feng X, Guo Z, Nourbakhsh M, Hauser H, Ganster R, Shao L et al. (2002). Proc Natl Acad Sci USA 99: 14212–14217.

  • Fischman TO, Hruza A, Da Niu X, Fossetta JD, Lunn CA, Dolphin E et al. (1999). Nat Struct Biol 6: 233–242.

  • Giepmans BN, Hengeveld T, Postma FR, Moolenaar WH . (2001). J Biol Chem 276: 8544–8549.

  • Hanke JH, Gardner JP, Sow RL, Changelian PS, Brissette WH, Weringer EJ et al. (1996). J Biol Chem 271: 695–701.

  • Huyer G, Liu S, Kelly J, Moffat J, Payette P, Kennedy B et al. (1997). J Biol Chem 272: 843–851.

  • Irby RB, Yeatman TJ . (2000). Oncogene 19: 5636–5642.

  • Keely SJ, Calandrella SO, Barrett KE . (2000). J Biol Chem 275: 12619–12625.

  • Kimura H, Miura S, Shigematsu T, Ohkubo N, Tsuzuki Y, Kurose I et al. (1997). Dig Dis Sci 42: 1047–1054.

  • Kolodziejski PJ, Musial A, Koo JS, Eissa NT . (2002). Proc Natl Acad Sci USA 99: 12315–12320.

  • Kroncke KD, Fehsel K, Kolb-Bachofen V . (1998). Clin Exp Immunol 113: 147–156.

  • Latour S, Veillette A . (2001). Curr Opin Immunol 13: 299–306.

  • Lee J, Ryu H, Ferrante RJ, Morris Jr SM, Ratan RR . (2003). Proc Natl Acad Sci USA 100: 4843–4848.

  • MacMicking J, Xie QW, Nathan C . (1997). Annu Rev Immunol 15: 323–350.

  • Mariner DJ, Anastasiadis P, Keilhack H, Bohmer FD, Wang J, Reynolds AB . (2001). J Biol Chem 276: 28006–28013.

  • Miller MJS, Sandoval M . (1999). Am J Physiol 276: G795–G799.

  • Musial A, Eissa NT . (2001). J Biol Chem 276: 24268–24273.

  • Navarro-Lerida I, Corvi MM, Barrientos AA, Gavilanes F, Berthiaume LG, Rodriguez-Crespo I . (2004). J Biol Chem 279: 55682–55689.

  • Paccani SR, Patrussi L, Ulivieri C, Masferrer JL, D'Elios MM, Baldari CT . (2005). Blood 105: 2042–2048.

  • Pan J, Burgher KL, Szczepanik AM, Ringheim GE . (1996). Biochem J 314: 889–894.

  • Posner B, Faure R, Burgess J, Bevan A, Lachance D, Sun G et al. (1994). J Biol Chem 269: 4596–4604.

  • Rachmilewitz D, Stamler JS, Bachwich D, Karmeli F, Ackerman Z, Podolski DK . (1995). Gut 36: 718–723.

  • Ratovitski EA, Alam MR, Quick RA, McMillan A, Bao C, Kozlovsy C et al. (1999a). J Biol Chem 274: 993–999.

  • Ratovitski EA, Bao C, Quick RA, McMillan A, Kozlovsky C, Lowenstein CJ . (1999b). J Biol Chem 274: 30250–30257.

  • Rumbo M, Courjault-Gautier F, Sierro F, Sirard JC, Felley-Bosco E . (2005). FEBS J 272: 444–453.

  • Salh B, Wagey R, Marotta A, Tao JS, Pelech S . (1998). J Immunol 161: 6947–6954.

  • Sayle RA, Milner-White EJ . (1995). Trends Biochem Sci 20: 374.

  • Singer II, Kawka DW, Scott S, Weidner JR, Mumford RA, Riehl TE et al. (1996). Gastroenterology 111: 871–885.

  • Sondermann H, Kuriyan J . (2005). Cell 121: 158–160.

  • Songyang Z, Cantley LC . (1995). Trends Biochem Sci 20: 470–475.

  • Taylor BS, deVera ME, Ganster RW, Wang Q, Shapiro RA, Morris SM et al. (1998). J Biol Chem 273: 15148–15156.

  • Thomsen LL, Miles DW, Happerfield L, Bobrow LG, Knowles RG, Moncada S . (1995). Br J Cancer 72: 41–44.

  • Vecchini F, Pringault E, Billiar TR, Geller DA, Hausel P, Felley-Bosco E . (1997). Cell Growth Differ 8: 261–268.

  • Vodovotz Y, Russell D, Xie Q, Bogdan C, Nathan C . (1995). J Immunol 154: 2914–2925.

Download references


We thank J Dall'Aglio for his skillful assistance, Drs JR Weidner, W Moolenaar and C Baldari for their generous gift of the anti-human iNOS antiserum and Src or Fyn encoding plasmids, Drs B Hemmings and D Diviani for suggestions and Drs B Thorens, M vanBemmelen and D Firsov for critical reading of the manuscript. This work was supported by the Swiss National Science Foundation (SNSF 3100A0-103928) and EC Grant QLRT2001-02357.

Author information

Authors and Affiliations


Corresponding author

Correspondence to E Felley-Bosco.

Additional information

Supplementary Information accompanies the paper on Oncogene website (

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Hausel, P., Latado, H., Courjault-Gautier, F. et al. Src-mediated phosphorylation regulates subcellular distribution and activity of human inducible nitric oxide synthase. Oncogene 25, 198–206 (2006).

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI:


  • inducible nitric oxide synthase
  • Src kinase
  • tyrosine phosphorylation


Quick links