Fig. 2.
Expression of cdka;1-as largely restores the defects of cdka;1 mutants. a Wild type rosette plants, approximately 1 month after sowing. Scale bar: 1 cm. b The cdka;1 homozygous mutants are extremely dwarf and can only grow on agar or in liquid media due to the absence of a functional root. Ruler scale is at cm. c Scanning electron micrograph of a 3 month-old cdka;1 homozygous mutant plant seen in b. Scale bar: 1 mm. d The expression of the cdka;1-as (CDKA;1 F80G) mutant largely rescues the development of homozygous cdka;1 null mutants that develop a root and can grow on soil. Plant shown was planted the same time as the wild-type control in panel a. Scale bar: 1 cm. e The expression of cdka;1-as does not confer a gain-of-function effect as seen in plants that contain the as allele in the heterozygous cdka;1 mutant background. Plant shown was planted the same time as the wild-type control in panel a. Scale bar: 1 cm. f Heterozygous cdka;1 mutants as a control. Plant shown was planted the same time as the wild-type control in panel a. Scale bar: 1 cm. g p13Suc1-associated protein kinase activity purified from wild-type plants (WT), cdka;1-as plants (in homozygous cdka;1 −/− (−/−), and heterozygous cdka;1 +/− (+/−) mutant background) or buffer (Mock), respectively, against bovine histone H1 as a generic substrate. Proteins were subjected to SDS-PAGE after the kinase reaction and stained with Coomassie brilliant blue R-250 demonstrating equal loading of the substrate. Abbreviations: p-H1 for radio-labeled histone H1 resulting from kinase assays with radio-labeled ATP, H1 for histone H1. h Protein blot analysis extracts from the wildtype plant (left), cdka;1-as plant in cdka;1 −/− (middle) and cdka;1 +/− (right) background, respectively, were probed with the antibody raised against the PSTAIRE cyclin-binding motif demonstrating comparable level of CDKA;1 in the indicated genotypes