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Published on July 25, 2008; 10.1104/pp.108.121897

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Received April 23, 2008
Accepted July 14, 2008

In planta analysis of the cell cycle-dependent localization of AtCDC48A and its critical roles in cell division, expansion, and differentiation

Sookhee Park , David Michael Rancour *, and Sebastian York Bednarek *

Department of Biochemistry, University of Wisconsin, 433 Babcock Dr., Madison, WI 53706 U.S.A.; Program in Cellular and Molecular Biology, University of Wisconsin, Madison, WI 53706 U.S.A.

* Corresponding author; email: sybednar{at}wisc.edu; drancour@wisc.edu.

CDC48/p97 is a conserved homohexameric AAA-ATPase chaperone required for a variety of cellular processes but whose role in the development of a multicellular model system has not been examined. Here we have used reverse genetics, visualization of a functional Arabidopsis CDC48 fluorescent fusion protein, and morphological analysis to examine the subcellular distribution and requirements for AtCDC48A in planta. Homozygous Atcdc48A T-DNA insertion mutants arrest during seedling development exhibiting decreased cell expansion and display pleiotropic defects in pollen and embryo development. Atcdc48A insertion alleles show significantly reduced male transmission efficiency due to defects in pollen tube growth. YFP-AtCDC48A, a fusion protein that functionally complements the insertion mutant defects, localizes in the nucleus, cytoplasm, and is recruited to the division midzone during cytokinesis. The pattern of nuclear localization differs according to the stage of the cell cycle and differentiation state. Inducible expression of an Atcdc48A Walker A ATPase mutant in planta results in cytokinesis abnormalities, aberrant cell divisions, and root trichoblast differentiation defects apparent in excessive root hair emergence. At the biochemical level, our data suggests that the endogenous steady-state protein level of AtCDC48A is dependent upon the presence of ATPase-active AtCDC48A. These results demonstrate that CDC48A/p97 is critical for cytokinesis, cell expansion and differentiation in plants.




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