In planta analysis of the cell cycle-dependent localization of AtCDC48A and its critical roles in cell division, expansion, and differentiation.
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 (Arabidopsis thaliana) 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 displaying pleiotropic defects in pollen and embryo development. Atcdc48A insertion alleles show significantly reduced male transmission efficiency due to defects in pollen tube growth. Yellow fluorescent protein-AtCDC48A, a fusion protein that functionally complements the insertion mutant defects, localizes in the nucleus and cytoplasm and is recruited to the division mid-zone 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 suggest 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.
Park
Sookhee
S
Department of Biochemistry , University of Wisconsin, Madison, Wisconsin 53706, USA.
Rancour
David Michael
DM
Bednarek
Sebastian York
SY
D000074183:ATPases Associated with Diverse Cellular Activities; D017360:Arabidopsis / Q000166:cytology / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D018797:Cell Cycle Proteins / Q000378:metabolism*; D002454:Cell Differentiation; D049108:Cell Enlargement; D048749:Cytokinesis; D006720:Homozygote; D016254:Mutagenesis, Insertional; D010641:Phenotype; D018517:Plant Roots / Q000166:cytology / Q000254:growth & development; D030821:Plants, Genetically Modified; D011058:Pollen / Q000254:growth & development; D011401:Promoter Regions, Genetic; D036226:Seedlings / Q000378:metabolism
D029681:Arabidopsis Proteins; D018797:Cell Cycle Proteins; D000074183:ATPases Associated with Diverse Cellular Activities; C525275:CDC48A protein, Arabidopsis