γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis.
γ-Tubulin is the key protein for microtubule nucleation. Duplication of the γ-tubulin gene occurred several times during evolution, and in mammals γ-tubulin genes encode proteins which share ∼97% sequence identity. Previous analysis of Tubg1 and Tubg2 knock-out mice has suggested that γ-tubulins are...
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2012
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oai:doaj.org-article:6f2c2d96bd124581b6fc0bf4cba76dc42021-11-18T07:31:02Zγ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis.1932-620310.1371/journal.pone.0029919https://doaj.org/article/6f2c2d96bd124581b6fc0bf4cba76dc42012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22235350/?tool=EBIhttps://doaj.org/toc/1932-6203γ-Tubulin is the key protein for microtubule nucleation. Duplication of the γ-tubulin gene occurred several times during evolution, and in mammals γ-tubulin genes encode proteins which share ∼97% sequence identity. Previous analysis of Tubg1 and Tubg2 knock-out mice has suggested that γ-tubulins are not functionally equivalent. Tubg1 knock-out mice died at the blastocyst stage, whereas Tubg2 knock-out mice developed normally and were fertile. It was proposed that γ-tubulin 1 represents ubiquitous γ-tubulin, while γ-tubulin 2 may have some specific functions and cannot substitute for γ-tubulin 1 deficiency in blastocysts. The molecular basis of the suggested functional difference between γ-tubulins remains unknown. Here we show that exogenous γ-tubulin 2 is targeted to centrosomes and interacts with γ-tubulin complex proteins 2 and 4. Depletion of γ-tubulin 1 by RNAi in U2OS cells causes impaired microtubule nucleation and metaphase arrest. Wild-type phenotype in γ-tubulin 1-depleted cells is restored by expression of exogenous mouse or human γ-tubulin 2. Further, we show at both mRNA and protein levels using RT-qPCR and 2D-PAGE, respectively, that in contrast to Tubg1, the Tubg2 expression is dramatically reduced in mouse blastocysts. This indicates that γ-tubulin 2 cannot rescue γ-tubulin 1 deficiency in knock-out blastocysts, owing to its very low amount. The combined data suggest that γ-tubulin 2 is able to nucleate microtubules and substitute for γ-tubulin 1. We propose that mammalian γ-tubulins are functionally redundant with respect to the nucleation activity.Stanislav VinopalMarkéta CernohorskáVadym SulimenkoTetyana SulimenkoVěra VoseckáMatyáš FlemrEduarda DráberováPavel DráberPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 1, p e29919 (2012) |
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Medicine R Science Q Stanislav Vinopal Markéta Cernohorská Vadym Sulimenko Tetyana Sulimenko Věra Vosecká Matyáš Flemr Eduarda Dráberová Pavel Dráber γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| description |
γ-Tubulin is the key protein for microtubule nucleation. Duplication of the γ-tubulin gene occurred several times during evolution, and in mammals γ-tubulin genes encode proteins which share ∼97% sequence identity. Previous analysis of Tubg1 and Tubg2 knock-out mice has suggested that γ-tubulins are not functionally equivalent. Tubg1 knock-out mice died at the blastocyst stage, whereas Tubg2 knock-out mice developed normally and were fertile. It was proposed that γ-tubulin 1 represents ubiquitous γ-tubulin, while γ-tubulin 2 may have some specific functions and cannot substitute for γ-tubulin 1 deficiency in blastocysts. The molecular basis of the suggested functional difference between γ-tubulins remains unknown. Here we show that exogenous γ-tubulin 2 is targeted to centrosomes and interacts with γ-tubulin complex proteins 2 and 4. Depletion of γ-tubulin 1 by RNAi in U2OS cells causes impaired microtubule nucleation and metaphase arrest. Wild-type phenotype in γ-tubulin 1-depleted cells is restored by expression of exogenous mouse or human γ-tubulin 2. Further, we show at both mRNA and protein levels using RT-qPCR and 2D-PAGE, respectively, that in contrast to Tubg1, the Tubg2 expression is dramatically reduced in mouse blastocysts. This indicates that γ-tubulin 2 cannot rescue γ-tubulin 1 deficiency in knock-out blastocysts, owing to its very low amount. The combined data suggest that γ-tubulin 2 is able to nucleate microtubules and substitute for γ-tubulin 1. We propose that mammalian γ-tubulins are functionally redundant with respect to the nucleation activity. |
| format |
article |
| author |
Stanislav Vinopal Markéta Cernohorská Vadym Sulimenko Tetyana Sulimenko Věra Vosecká Matyáš Flemr Eduarda Dráberová Pavel Dráber |
| author_facet |
Stanislav Vinopal Markéta Cernohorská Vadym Sulimenko Tetyana Sulimenko Věra Vosecká Matyáš Flemr Eduarda Dráberová Pavel Dráber |
| author_sort |
Stanislav Vinopal |
| title |
γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| title_short |
γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| title_full |
γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| title_fullStr |
γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| title_full_unstemmed |
γ-Tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| title_sort |
γ-tubulin 2 nucleates microtubules and is downregulated in mouse early embryogenesis. |
| publisher |
Public Library of Science (PLoS) |
| publishDate |
2012 |
| url |
https://doaj.org/article/6f2c2d96bd124581b6fc0bf4cba76dc4 |
| work_keys_str_mv |
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| _version_ |
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