A kinesin-like protein, KatAp, in the cells of arabidopsis and other plants

Bo Liu, Richard Cyr, Barry A. Palevitz

Research output: Contribution to journalArticle

108 Citations (Scopus)

Abstract

The kinesin-like proteins (KLPs) are a large family of plus- or minus-end-directed microtubule motors important in intracellular transport, mitosis, meiosis, and development. However, relatively little is known about plant KLPs. We prepared an antibody against two peptides in the microtubule binding domain of an Arabidopsis KLP (KatAp) encoded by the KatA gene, one of a family of genes encoding KLPs whose motor domain is located near the C terminus of the polypeptide. Such KLPs typically move materials toward the minus end of microtubules. An immunoreactive band (Mr of 140,000) corresponding to KatAp was demonstrated with this antibody on immunoblots of Arabidopsis seedling extracts. During immunofluorescence localizations, the antibody produced weak, variable staining in the cytoplasm and nucleus of interphase Arabidopsis suspension cells but much stronger staining of the mitotic apparatus during division. Staining was concentrated near the midzone during metaphase and was retained there during anaphase. The phragmoplast was also stained. Similar localization patterns were seen in tobacco BY-2 cells. The antibody produced a single band (Mr of 130,000) in murine brain fractions prepared according to procedures that enrich for KLPs (binding to microtubules in the presence of AMP-PNP but not ATP). A similar traction from carrot suspension cells yielded a cross-reacting polypeptide of similar apparent molecular mass. When dividing BY-2 cells were lysed in the presence of taxol and ATP, antibody staining moved rapidly toward the poles, supporting the presence of a minus-end motor. Movement did not occur without ATP, with AMP-PNP, or with ATP plus antibody. Our results indicate that the protein encoded by KatA, KatAp, is expressed in Arabidopsis and is specifically localized to the midzone of the mitotic apparatus and phragmoplast. A similar protein is also present in other species.

Original languageEnglish (US)
Pages (from-to)119-132
Number of pages14
JournalPlant Cell
Volume8
Issue number1
StatePublished - Jan 1 1996

Fingerprint

Kinesin
kinesin
Arabidopsis
Microtubules
microtubules
antibodies
Antibodies
Adenosine Triphosphate
Adenylyl Imidodiphosphate
Proteins
Staining and Labeling
proteins
cells
mitotic spindle apparatus
Spindle Apparatus
cell suspension culture
polypeptides
Suspensions
Peptides
Anaphase

All Science Journal Classification (ASJC) codes

  • Plant Science
  • Cell Biology

Cite this

Liu, B., Cyr, R., & Palevitz, B. A. (1996). A kinesin-like protein, KatAp, in the cells of arabidopsis and other plants. Plant Cell, 8(1), 119-132.
Liu, Bo ; Cyr, Richard ; Palevitz, Barry A. / A kinesin-like protein, KatAp, in the cells of arabidopsis and other plants. In: Plant Cell. 1996 ; Vol. 8, No. 1. pp. 119-132.
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Liu, B, Cyr, R & Palevitz, BA 1996, 'A kinesin-like protein, KatAp, in the cells of arabidopsis and other plants', Plant Cell, vol. 8, no. 1, pp. 119-132.

A kinesin-like protein, KatAp, in the cells of arabidopsis and other plants. / Liu, Bo; Cyr, Richard; Palevitz, Barry A.

In: Plant Cell, Vol. 8, No. 1, 01.01.1996, p. 119-132.

Research output: Contribution to journalArticle

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N2 - The kinesin-like proteins (KLPs) are a large family of plus- or minus-end-directed microtubule motors important in intracellular transport, mitosis, meiosis, and development. However, relatively little is known about plant KLPs. We prepared an antibody against two peptides in the microtubule binding domain of an Arabidopsis KLP (KatAp) encoded by the KatA gene, one of a family of genes encoding KLPs whose motor domain is located near the C terminus of the polypeptide. Such KLPs typically move materials toward the minus end of microtubules. An immunoreactive band (Mr of 140,000) corresponding to KatAp was demonstrated with this antibody on immunoblots of Arabidopsis seedling extracts. During immunofluorescence localizations, the antibody produced weak, variable staining in the cytoplasm and nucleus of interphase Arabidopsis suspension cells but much stronger staining of the mitotic apparatus during division. Staining was concentrated near the midzone during metaphase and was retained there during anaphase. The phragmoplast was also stained. Similar localization patterns were seen in tobacco BY-2 cells. The antibody produced a single band (Mr of 130,000) in murine brain fractions prepared according to procedures that enrich for KLPs (binding to microtubules in the presence of AMP-PNP but not ATP). A similar traction from carrot suspension cells yielded a cross-reacting polypeptide of similar apparent molecular mass. When dividing BY-2 cells were lysed in the presence of taxol and ATP, antibody staining moved rapidly toward the poles, supporting the presence of a minus-end motor. Movement did not occur without ATP, with AMP-PNP, or with ATP plus antibody. Our results indicate that the protein encoded by KatA, KatAp, is expressed in Arabidopsis and is specifically localized to the midzone of the mitotic apparatus and phragmoplast. A similar protein is also present in other species.

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