TY - JOUR
T1 - Polyglycylation domain of β-tubulin maintains axonemal architecture and affects cytokinesis in Tetrahymena
AU - Thazhath, Rupal
AU - Liu, Chengbao
AU - Gaertig, Jacek
N1 - Funding Information:
ACKNOWLEDGEMENTS This work was supported by a National Institutes of Health grant GM 54017 and an American Cancer Society grant RPG-99-245-01-CSM. We are grateful to the staff of the Center for Advanced Ultrastructural Research at the University of Georgia for training R.T. and for extensive technical assistance. We thank J. Frankel and J. Rosenbaum for numerous comments and stimulating ideas. We also thank M. Gorovsky, M. Jerka-Dziadosz, N. Williams and J. Lauderdale for critical reading of the manuscript, J. Salisbury for the 20H5 antibody, G. Piperno for the 6-11 B-1 antibody, and M.-H. Bré and N. Levilliers for the AXO49 antibodies. The 12G10 monoclonal antibody was produced by E.M. Nelsen and J. Frankel, and will be available from the Developmental Studies Hybridoma Bank developed under the auspices of the NICHD and maintained by the University of Iowa Department of Biological Sciences. Correspondence and requests for material should be addressed to J.G.
PY - 2002
Y1 - 2002
N2 - Polyglycylation occurs through the post-translational addition of a polyglycine peptide to the γ-carboxyl group of glutamic acids near the C terminus of α- and β-tubulin1, and has been found only in cells with axonemes, from protists to humans2,3. In Tetrahymena thermophila, multiple sites of polyglycylation on α-tubulin are dispensable. By contrast, mutating similar sites on β-tubulin has site-specific effects, affecting cell motility and cytokinesis, or resulting in cell death4. Here, we address the lethality of a polyglycylation deficiency in T. thermophila using heterokaryons5. Cells with a lethal mutation in the polyglycylation domain of β-tubulin assembled axonemes that lack the central pair, B-subfibres and the transitional zone of outer microtubules (MTs). Furthermore, an arrest in cytokinesis occurred, and was associated with incomplete severing of cortical MTs positioned near the cleavage furrow. Thus, tubulin polyglycylation is required for the maintenance of some stable microtubular organelles that are all known to be polyglycylated in vivo, but its effects on MTs appear to be organelle-specific.
AB - Polyglycylation occurs through the post-translational addition of a polyglycine peptide to the γ-carboxyl group of glutamic acids near the C terminus of α- and β-tubulin1, and has been found only in cells with axonemes, from protists to humans2,3. In Tetrahymena thermophila, multiple sites of polyglycylation on α-tubulin are dispensable. By contrast, mutating similar sites on β-tubulin has site-specific effects, affecting cell motility and cytokinesis, or resulting in cell death4. Here, we address the lethality of a polyglycylation deficiency in T. thermophila using heterokaryons5. Cells with a lethal mutation in the polyglycylation domain of β-tubulin assembled axonemes that lack the central pair, B-subfibres and the transitional zone of outer microtubules (MTs). Furthermore, an arrest in cytokinesis occurred, and was associated with incomplete severing of cortical MTs positioned near the cleavage furrow. Thus, tubulin polyglycylation is required for the maintenance of some stable microtubular organelles that are all known to be polyglycylated in vivo, but its effects on MTs appear to be organelle-specific.
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U2 - 10.1038/ncb764
DO - 10.1038/ncb764
M3 - Article
C2 - 11862218
AN - SCOPUS:0036125515
SN - 1465-7392
VL - 4
SP - 256
EP - 259
JO - Nature cell biology
JF - Nature cell biology
IS - 3
ER -