Dynamic interaction of NtMAP65-1a with microtubules in vivo

HY Chang, AP Smertenko, H Igarashi, DP Dixon, PJ Hussey*

*Corresponding author for this work

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Plant microtubules are intrinsically more dynamic than those from animals. We know little about the dynamics of the interaction of plant microtubule-associated proteins (MAPs) with microtubules. Here, we have used tobacco and Arabidopsis MAPs with relative molecular mass 65 kDa (NtMAP65-1a and AtMAP65-1), to study their interaction with microtubules in vivo. Using fluorescence recovery after photobleaching we report that the turnover of both NtMAP65-1a and AtMAP65-1 bound to microtubules is four- to fivefold faster than microtubule treadmilling (13 seconds compared with 56 seconds, respectively) and that the replacement of NtMAP65-1a on microtubules is by random association rather than by translocation along microtubules. MAP65 will only bind polymerised microtubules and not its component tubulin dimers. The turnover of NtMAP65-1a and AtMAP65-1 on microtubules is similar in the interphase cortical array, the preprophase band and the phragmoplast, strongly suggesting that their role in these arrays is the same. NtMAP65-1a and AtMAP65-1 are not observed to bind microtubules in the metaphase spindle and their rate of recovery is consistent with their cytoplasmic localisation. In addition, the dramatic reappearance of NtMAP65-1a on microtubules at the spindle midzone in anaphase B suggests that NtMAP65-1a is controlled post-translationally. We conclude that the dynamic properties of these MAPs in vivo taken together with the fact that they have been shown not to effect microtubule polymerisation in vitro, makes them ideally suited to a role in crossbridging microtubules that need to retain spatial organisation in rapidly reorganising microtubule arrays.

Original languageEnglish
Pages (from-to)3195-3201
Number of pages7
JournalJournal of Cell Science
Volume118
Issue number14
DOIs
Publication statusPublished - 15 Jul 2005

Keywords

  • BINDING
  • PROTEIN
  • INSTABILITY
  • FRAP
  • CELLS
  • microtubule-associated protein
  • microtubules
  • GFP
  • MAP65
  • CYTOSKELETON
  • CYTOKINETIC PHRAGMOPLAST
  • PLANTS
  • ARABIDOPSIS-THALIANA

Cite this

Chang, HY., Smertenko, AP., Igarashi, H., Dixon, DP., & Hussey, PJ. (2005). Dynamic interaction of NtMAP65-1a with microtubules in vivo. Journal of Cell Science, 118(14), 3195-3201. https://doi.org/10.1242/jcs.02433
Chang, HY ; Smertenko, AP ; Igarashi, H ; Dixon, DP ; Hussey, PJ. / Dynamic interaction of NtMAP65-1a with microtubules in vivo. In: Journal of Cell Science. 2005 ; Vol. 118, No. 14. pp. 3195-3201.
@article{4203c1ef80bf4c4ab31767ff8b2fc83a,
title = "Dynamic interaction of NtMAP65-1a with microtubules in vivo",
abstract = "Plant microtubules are intrinsically more dynamic than those from animals. We know little about the dynamics of the interaction of plant microtubule-associated proteins (MAPs) with microtubules. Here, we have used tobacco and Arabidopsis MAPs with relative molecular mass 65 kDa (NtMAP65-1a and AtMAP65-1), to study their interaction with microtubules in vivo. Using fluorescence recovery after photobleaching we report that the turnover of both NtMAP65-1a and AtMAP65-1 bound to microtubules is four- to fivefold faster than microtubule treadmilling (13 seconds compared with 56 seconds, respectively) and that the replacement of NtMAP65-1a on microtubules is by random association rather than by translocation along microtubules. MAP65 will only bind polymerised microtubules and not its component tubulin dimers. The turnover of NtMAP65-1a and AtMAP65-1 on microtubules is similar in the interphase cortical array, the preprophase band and the phragmoplast, strongly suggesting that their role in these arrays is the same. NtMAP65-1a and AtMAP65-1 are not observed to bind microtubules in the metaphase spindle and their rate of recovery is consistent with their cytoplasmic localisation. In addition, the dramatic reappearance of NtMAP65-1a on microtubules at the spindle midzone in anaphase B suggests that NtMAP65-1a is controlled post-translationally. We conclude that the dynamic properties of these MAPs in vivo taken together with the fact that they have been shown not to effect microtubule polymerisation in vitro, makes them ideally suited to a role in crossbridging microtubules that need to retain spatial organisation in rapidly reorganising microtubule arrays.",
keywords = "BINDING, PROTEIN, INSTABILITY, FRAP, CELLS, microtubule-associated protein, microtubules, GFP, MAP65, CYTOSKELETON, CYTOKINETIC PHRAGMOPLAST, PLANTS, ARABIDOPSIS-THALIANA",
author = "HY Chang and AP Smertenko and H Igarashi and DP Dixon and PJ Hussey",
year = "2005",
month = "7",
day = "15",
doi = "10.1242/jcs.02433",
language = "English",
volume = "118",
pages = "3195--3201",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "14",

}

Chang, HY, Smertenko, AP, Igarashi, H, Dixon, DP & Hussey, PJ 2005, 'Dynamic interaction of NtMAP65-1a with microtubules in vivo', Journal of Cell Science, vol. 118, no. 14, pp. 3195-3201. https://doi.org/10.1242/jcs.02433

Dynamic interaction of NtMAP65-1a with microtubules in vivo. / Chang, HY; Smertenko, AP; Igarashi, H; Dixon, DP; Hussey, PJ.

In: Journal of Cell Science, Vol. 118, No. 14, 15.07.2005, p. 3195-3201.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Dynamic interaction of NtMAP65-1a with microtubules in vivo

AU - Chang, HY

AU - Smertenko, AP

AU - Igarashi, H

AU - Dixon, DP

AU - Hussey, PJ

PY - 2005/7/15

Y1 - 2005/7/15

N2 - Plant microtubules are intrinsically more dynamic than those from animals. We know little about the dynamics of the interaction of plant microtubule-associated proteins (MAPs) with microtubules. Here, we have used tobacco and Arabidopsis MAPs with relative molecular mass 65 kDa (NtMAP65-1a and AtMAP65-1), to study their interaction with microtubules in vivo. Using fluorescence recovery after photobleaching we report that the turnover of both NtMAP65-1a and AtMAP65-1 bound to microtubules is four- to fivefold faster than microtubule treadmilling (13 seconds compared with 56 seconds, respectively) and that the replacement of NtMAP65-1a on microtubules is by random association rather than by translocation along microtubules. MAP65 will only bind polymerised microtubules and not its component tubulin dimers. The turnover of NtMAP65-1a and AtMAP65-1 on microtubules is similar in the interphase cortical array, the preprophase band and the phragmoplast, strongly suggesting that their role in these arrays is the same. NtMAP65-1a and AtMAP65-1 are not observed to bind microtubules in the metaphase spindle and their rate of recovery is consistent with their cytoplasmic localisation. In addition, the dramatic reappearance of NtMAP65-1a on microtubules at the spindle midzone in anaphase B suggests that NtMAP65-1a is controlled post-translationally. We conclude that the dynamic properties of these MAPs in vivo taken together with the fact that they have been shown not to effect microtubule polymerisation in vitro, makes them ideally suited to a role in crossbridging microtubules that need to retain spatial organisation in rapidly reorganising microtubule arrays.

AB - Plant microtubules are intrinsically more dynamic than those from animals. We know little about the dynamics of the interaction of plant microtubule-associated proteins (MAPs) with microtubules. Here, we have used tobacco and Arabidopsis MAPs with relative molecular mass 65 kDa (NtMAP65-1a and AtMAP65-1), to study their interaction with microtubules in vivo. Using fluorescence recovery after photobleaching we report that the turnover of both NtMAP65-1a and AtMAP65-1 bound to microtubules is four- to fivefold faster than microtubule treadmilling (13 seconds compared with 56 seconds, respectively) and that the replacement of NtMAP65-1a on microtubules is by random association rather than by translocation along microtubules. MAP65 will only bind polymerised microtubules and not its component tubulin dimers. The turnover of NtMAP65-1a and AtMAP65-1 on microtubules is similar in the interphase cortical array, the preprophase band and the phragmoplast, strongly suggesting that their role in these arrays is the same. NtMAP65-1a and AtMAP65-1 are not observed to bind microtubules in the metaphase spindle and their rate of recovery is consistent with their cytoplasmic localisation. In addition, the dramatic reappearance of NtMAP65-1a on microtubules at the spindle midzone in anaphase B suggests that NtMAP65-1a is controlled post-translationally. We conclude that the dynamic properties of these MAPs in vivo taken together with the fact that they have been shown not to effect microtubule polymerisation in vitro, makes them ideally suited to a role in crossbridging microtubules that need to retain spatial organisation in rapidly reorganising microtubule arrays.

KW - BINDING

KW - PROTEIN

KW - INSTABILITY

KW - FRAP

KW - CELLS

KW - microtubule-associated protein

KW - microtubules

KW - GFP

KW - MAP65

KW - CYTOSKELETON

KW - CYTOKINETIC PHRAGMOPLAST

KW - PLANTS

KW - ARABIDOPSIS-THALIANA

U2 - 10.1242/jcs.02433

DO - 10.1242/jcs.02433

M3 - Article

VL - 118

SP - 3195

EP - 3201

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 14

ER -

Chang HY, Smertenko AP, Igarashi H, Dixon DP, Hussey PJ. Dynamic interaction of NtMAP65-1a with microtubules in vivo. Journal of Cell Science. 2005 Jul 15;118(14):3195-3201. https://doi.org/10.1242/jcs.02433