Proliferating Cell Nuclear Antigen(TaPCNA), a Pyruvate Dehydrogenase Kinase(TaPDK)-interacting Protein, Is Involved in the Regulation of Microspore Cell Cycle in Wheat(Triticum aestivum)
Abstract：It has been documented that pyruvate dehydrogenase kinase(PDK) is involved in the regulation of pyruvate dehydrogenase complex activity in mitochondria. However, very little is known about the regulatory mechanism of PDK. Our previous study showed that there was difference in the expression of TaPDK between genetic sterile line and physiological sterile line. To further study the acting factor regulating TaPDK during anther abortion induced by SQ-1, the interaction protein of TaPDK was screened via yeast two-hybrid technique. The yeast (Saccharomyces cerevisiae) strain Y2HGold containing pGBKT7-PDK plasmid and, library yeast strain Y187 were cultured by yeast mating. Then, the mated culture was incubated on SD/-Ade/-His/-Leu/-Trp plate before selection of clones with diameter larger than 2 mm, and further incubated on SD/-Ade/-His/-Leu/-Trp/AbA/X-α-Gal plate for screening blue clones. A new binding protein proliferating cell nuclear antigen was obtained, and was designated as proliferating cell nuclear antigen(TaPCNA). The results from pollen development and quantitative expression indicated that the expression of TaPCNA was scarcely expressed in leaf, and mainly expressed in different stage of pollen development. Especially, its transcripts accumulated predominantly in physiological sterile line at bicellular and tricellular stage, which suggested that TaPCNA was closely involved with the cell cycle of microspore development, and provided the basic date for further study of the abnormal cell cycle of microspore development in physiological sterile line.
