cDNA isolation, characterization, and protein intracellular localization of a katanin-like p60 subunit from Arabidopsis thaliana |
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Authors: | R S McClinton J S Chandler J Callis |
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Institution: | (1) Department of Biology, University of Louisiana at Lafayette, Lafayette, Louisiana;(2) Section of Molecular and Cellular Biology, University of California-Davis, Davis, California;(3) Department of Biology, University of Louisiana at Lafayette, P.O. Box 42451, 70504-2451 Lafayette, LA, USA |
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Abstract: | Summary Katanin, a heterodimeric protein with ATP-dependent microtubule-severing activity, localizes to the centrosome in animal cells.
Widespread occurrence is suspected as several species contain homologs to the katanin p60 subunit. Recently we isolated anArabidopsis thaliana cDNA with significant identity to the p60 subunit of sea urchin katanin. Like p60, the encoded protein is a member of the
AAA superfamily of ATPases, containing the Walker ATP binding consensus and the signature AAA minimal consensus sequences
within a single larger AAA/CAD amino acid motif. Phylogenetic analysis placed the encoded protein in the AAA subfamily of
cytoskeleton-interactive proteins, where it formed a strongly supported clade with 4 other members identified as katanin p60
subunits. The clone was named AtKSSArabidopsis thaliana kataninlike protein small subunit). Western blots, performed using a polyclonal antibody raised against recombinant AtKSS,
revealed AtKSS is present in protein extracts of all Arabidopsis organs examined. To evaluate potential interactions between
AtKSS and the cytoskeleton, the intracellular localization of AtKSS was correlated with that of tubulin. AtKSS was found in
perinuclear regions during interphase, surrounding the spindle poles during mitosis, but was absent from the preprophase band
and phragmoplast microtubule arrays. These data support the thesis that AtKSS is an Arabidopsis homolog of the p60 subunit
of katanin. Its cell cycle-dependent distribution is consistent with microtubule-severing activity, but additional studies
will better define its role. |
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Keywords: | Arabidopsis thaliana AAA protein ATPase Cytoskeleton Katanin |
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