Figure 4.

Complementation of fibroblasts from a CG-J Zellweger patient. (A) Partial sequence and deduced amino acid sequence of PEX19 cDNA isolated from a normal control (Left) and a ZS patient PBDJ-01 (Center) are shown. PCR was also done for DNA from PBDJ-01 fibroblasts (Right). One-base insertion A⁷⁶⁴ (shaded), in a codon for Met²⁵⁵, causes a frameshift in PBDJ-01 PEX19 sequence. (B) Sequence comparison of the C-terminal part of Pex19p each from a control and a patient PBDJ-01. Amino acid sequence resulted from the frameshift by the 1-bp insertion in PBDJ-01 is underlined. The arrowhead indicates the position of frameshift mutation. The CAAX box is double-underlined. (C) Transfection of PEX19 from a normal control and a CG-J patient (PBDJ-01). (a) PBDJ-01 fibroblasts. (b) PBDJ-01 fibroblasts were transfected with pUcD2Hyg⋅HsPEX19. (c and d) PBDJ-01 fibroblasts were transfected with PBDJ-01-derived PEX19 cDNA, PEX19A764ins, and a mutant PEX19, PEX19C296S, respectively. (e and f) flag-tagged PEX19C296S, flag-PEX19C296S, was expressed in ZP119. Cells were stained with antibodies to human catalase (a–d), rat catalase (e), and flag (f). Note that peroxisomes were restored only in b, but not in c–f. (Bar = 20 μm.)