Identification of Ppa1 and Ppa2 proteins. An immunoblot with anti-ppa2 antibody is shown (ref.1). lane 1: Wild-type S. pombe lane 2: ppa1 lane 3: ppa2 lane 4: Wild-type carrying a multicopy plasmid with ppa1 gene lane 5: Wild-type carrying a multicopy plasmid with ppa2 gene lane 6:Wild-type carrying a multicopy plasmid with ADH promoter ligated with ppa2 gene The positions of ppa1 (36 kDa) and ppa2 (39 kDa) polypeptide bands are indicated.

Cellular location of Ppa1 and Ppa2. Indirect immunofluorescence microscopy of ppa2 deletion, wild type (WT), and wild-type overexpressing ppa2 (nmt-ppa2, 18hr) was done, using anti-ppa2 antibody (right); The same cells stained by DAPI are also shown (left). Immunofluorescence was hardly detected in ppa2 cells, whereas cytoplasmic immunofluorescence was abundant in wild-type cells. Wild-type cells carrying nmt-ppa2 plasmid overexpress Ppa2 protein in the absence of thiamine for 18 hr. Immunofluorescence was enhanced further in the cytoplasm, often accumulated at the nuclear periphery or within restricted domains. The deformation of chromosomal DNA was also visible in overexpressed cells. Bar, 10um