tailieunhanh - Báo cáo Y học: Incorporation of 3-nitrotyrosine into the C-terminus of a-tubulin is reversible and not detrimental to dividing cells

The C-terminus of thea-chain of tubulin is subject to reversible incorporationof tyrosineby tubulin tyrosine ligase and removal by tubulin carboxypeptidase. Thus, microtu-bules rich in either tyrosinated or detyrosinated tubulin can coexist in the cell. Substitution of the terminal tyrosine by 3-nitrotyrosine has been claimed to cause microtubule dys-function and consequent injury of epithelial lung carcinoma A549 cells. Nitrotyrosine is formed in cells by nitration of tyrosine by nitric oxide-derived species | Eur. J. Biochem. 269 5037-5045 2002 FEBS 2002 doi Incorporation of 3-nitrotyrosine into the C-terminus of a-tubulin is reversible and not detrimental to dividing cells C. Gastein Bisig Silvia A. Purro Maria A. Contin Hector S. Barra and Carlos A. Arce Centro de Investigaciones en Quimica Biologica de Cordoba Departamento de Quimica Biologica Universidad Nacional de Cordoba Argentina The C-terminus of the a-chain of tubulin is subject to reversible incorporation of tyrosine by tubulin tyrosine ligase and removal by tubulin carboxypeptidase. Thus microtubules rich in either tyrosinated or detyrosinated tubulin can coexist in the cell. Substitution of the terminal tyrosine by 3-nitrotyrosine has been claimed to cause microtubule dysfunction and consequent injury of epithelial lung carcinoma A549 cells. Nitrotyrosine is formed in cells by nitration of tyrosine by nitric oxide-derived species. We studied properties of tubulin modified by in vitro nitrotyrosination at the C-terminus of the a-subunit and the consequences for cell functioning. Nitrotyrosinated tubulin was a good substrate of tubulin carboxypeptidase and showed a similar capability to assemble into microtubules in vitro to that of tyrosinated tubulin. Tubulin of C6 cells cultured in F12K medium in the presence of 500 M nitrotyrosine became fully nitrotyrosi-nated. This nitrotyrosination was shown to be reversible. No changes in morphology proliferation or viability were observed during cycles of nitrotyrosination denitrotyrosin-ation and re-nitrotyrosination. Similar results were obtained with CHO COS-7 HeLa NIH-3T3 NIH-3T3 TTL- and A549 cells. C6 and A549 cells were subjected to several passages during 45days or more in the continuous presence of 500 M nitrotyrosine without noticeable alteration of morphology viability or proliferation. The microtubular networks visualized by immunofluorescence with antibodies to nitrotyrosinated and total tubulin were identical. .

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