Cellular interactions and tubulin detyrosination in fibroblastic and epithelial cells
作者:
Marie‐Hélèn Bré,
Rainer Pepperkok,
Thomas E. Kreis,
Eric Karsenti,
期刊:
Biology of the Cell
(WILEY Available online 1991)
卷期:
Volume 71,
issue 1‐2
页码: 149-160
ISSN:0248-4900
年代: 1991
DOI:10.1016/0248-4900(91)90061-Q
出版商: Blackwell Publishing Ltd
关键词: microtubules;detyrosimation;cellular interactions;epithelial cells;MDCK
数据来源: WILEY
摘要:
Summary—In mammalian cells most microtubules are enriched in tyrosinated α‐tubulin (tyr‐tubulin). Other subclasses of microtubules are present in variable amounts and some are enriched in detyrosinated α‐tubulin (glu‐tubulin). We examined the effect of cell‐cell interactions on the level of glu‐tubulin in microtubules. This was studied by quantitative immunofluorescence using antibodies against tyr‐ and glu‐tubulin. We found that in cells which have established cell‐cell contacts, the ratio of glu‐/tyr‐tubulin is higher than in isolated cells. We also examined the effect of cell‐cell interactions on the glu‐/tyr‐tubulin ratio by using the antibody blocking method of Schulze and Kirschner [42]. Microtubules containing mainly tyr‐tubulin had been blocked first by a polyclonal antibody against tyr‐tubulin and several layers of secondary antibodies. The unblocked microtubules were then labeled by a monoclonal antibody against α‐tubulin. Since the coating efficiency of microtubules by the anti‐tyr tubulin depends on the amount of tyr‐tubulin in each microtubule, this procedure allows the visualization of microtubules enriched or depleted in tyr‐tubulin in specific domains of each cell. Microtubules were more extensively blocked in subconfluent than in confluent cells and preferentially at the periphery of the cytoplasm. In cells present at the margin of an artificial wound produced in a confluent monolayer, the amount of blocked microtubules increased slowly with time (between 2 and 4 h). These results are consistent with the hypothesis that cell‐cell contacts lead to increased tubulin detyrosina
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