Import of microinjected proteins bearing the SKL peroxisomal targeting sequence into the peroxisomes of a human fibroblast cell line: evidence that virtually all peroxisomes are import-competent.
Peroxisomes import virtually all of their membrane and matrix proteins post-translationally. It is presently unknown whether, in mammalian cells, their exists a pool of mature peroxisomes which have received their complement of proteins and are import-incompetent. Previous work has shown that fibroblasts are capable of importing microinjected peroxisomal proteins into peroxisomes. This report describes the import of a hybrid peroxisomal protein into virtually all peroxisomes of the microinjected cell. The peroxisomal import was uniform in both short and long incubations. Pretreatment of the cells with cycloheximide did not affect the import of the peroxisomal protein, nor was there any difference in the distribution of the imported protein. Sequential microinjection experiments demonstrated that peroxisomes that had imported luciferase were capable of importing another peroxisomal protein injected 24 hours later. These results suggest that, in fibroblasts, all peroxisomes have associated protein-import machinery; this evidence does not support the hypothesis that there exists a pool of import-incompetent peroxisomes.
Hill
P E
PE
Department of Anatomy and Cell Biology, McGill University, Montreal, Canada.
Walton
P A
PA
D002460:Cell Line; D003513:Cycloheximide / Q000494:pharmacology; D005347:Fibroblasts; D006801:Humans; D007700:Kinetics; D008830:Microbodies / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D008845:Microinjections; D009842:Oligopeptides / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism*; D012709:Serum Albumin / Q000008:administration & dosage / Q000032:analysis / Q000378:metabolism*; D013997:Time Factors
D009842:Oligopeptides; D012709:Serum Albumin; C088449:seryl-lysyl-leucine; D003513:Cycloheximide