Integration of different keratins into the same filament system after microinjection of mRNA for epidermal keratins into kidney epithelial cells.
We have isolated poly (A)+ RNA, highly enriched in keratin mRNA from bovine muzzle epidermis, and injected it into epithelial cells of a different type, i.e., cultured kidney epithelial cells of the same (MDBK) or taxonomically distant (PtK2) species. Both recipient cell lines contain keratin polypeptides that are different from those present in epidermal cells. Using keratin subtype-specific antibodies in immunofluorescence and immunoelectron microscopy, we show that foreign keratin mRNAs when injected into a different type of epithelial cell can recruit polyribosomes and are translated together with the keratin mRNAs of the host cell. Foreign epidermal keratins are excluded from vimentin filaments and other structures but readily coassemble with the endogenous keratins and appear to be integrated into the meshwork of the preexisting kidney-type keratin filaments. Our observations indicate that different sets of keratin polypeptides from the same or different species can coassemble in the living cell into a common filament system. Thus we have developed a procedure that allows experimental alteration of the intermediate filament cytoskeleton within living epithelial cells.
D000818:Animals; D002417:Cattle; D002460:Cell Line; D003599:Cytoskeleton / Q000378:metabolism* / Q000648:ultrastructure; D004848:Epithelium / Q000378:metabolism; D005455:Fluorescent Antibody Technique; D007633:Keratins / Q000235:genetics* / Q000378:metabolism; D007668:Kidney / Q000378:metabolism*; D008854:Microscopy, Electron; D009928:Organ Specificity; D011061:Poly A / Q000235:genetics; D012313:RNA / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D012867:Skin / Q000378:metabolism*
D012333:RNA, Messenger; D011061:Poly A; D012313:RNA; D007633:Keratins