A GFP-MAP4 reporter gene for visualizing cortical microtubule rearrangements in living epidermal cells

Jan Marc, Cheryl L. Granger, Jennifer Brincat, Deborah D. Fisher, Teh Hui Kao, Andrew G. McCubbin, Richard J. Cyr

Research output: Contribution to journalArticlepeer-review

301 Scopus citations

Abstract

Microtubules influence morphogenesis by forming distinct geometrical arrays in the cell cortex, which in turn affect the deposition of cellulose microfibrils. Although many chemical and physical factors affect microtubule orientation, it is unclear how cortical microtubules in elongating cells maintain their ordered transverse arrays and how they reorganize into new geometries. To visualize these reorientations in living cells, we constructed a microtubule reporter gene by fusing the microtubule binding domain of the mammalian microtubule-associated protein 4 (MAP4) gene with the green fluorescent protein (GFP) gene, and transient expression of the recombinant protein in epidermal cells of fava bean was induced. The reporter protein decorates microtubules in vivo and binds to microtubules in vitro. Confocal microscopy and time-course analysis of labeled cortical arrays along the outer epidermal wall revealed the lengthening, shortening, and movement of microtubules; localized microtubule reorientations; and global microtubule reorganizations. The global microtubule orientation in some cells fluctuates about the transverse axis and may be a result of a cyclic self-correcting mechanism to maintain a net transverse orientation during cellular elongation.

Original languageEnglish (US)
Pages (from-to)1927-1939
Number of pages13
JournalPlant Cell
Volume10
Issue number11
DOIs
StatePublished - 1998

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology

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