Selective synthesis of mRNA and proteins by human peripheral blood neutrophils.
Human peripheral blood polymorphonuclear neutrophils (PMN) have been considered to be capable of little if any protein biosynthesis. We evaluated the ability of PMN to synthesize both mRNA and proteins. Using in vitro [35S]methionine pulse-chase labeling of purified PMN, followed by immunoprecipitation of cell lysates with immobilized mAb and analysis by gel electrophoresis, PMN were shown to synthesize CR1, FcR, CR3 alpha-chain, MHC class I, and actin. In contrast, incorporation of [35S]methionine into either CR3 beta-chain or the secondary granule protein lactoferrin was not detected. Purification of mRNA from PMN and analysis by Northern blots demonstrated the presence in PMN of CR1, actin, and MHC class I transcripts. However, despite the apparent lack of CR3 beta-chain biosynthesis, specific beta-chain message was detectable in PMN RNA. Inhibition of mRNA synthesis in PMN with actinomycin D resulted in decreased synthesis of nascent CR1, FcR, MHC class I, and actin compared with control cells. Thus, PMN continue to transcribe and translate the genes for certain membrane and cytoskeletal proteins. In contrast, the lack of detectable synthesis of either lactoferrin or CR3 beta-chain suggested that biosynthesis in circulating PMN is selective.
D001798:Blood Proteins / Q000096:biosynthesis*; D002465:Cell Movement; D003166:Complement Activating Enzymes / Q000096:biosynthesis; D003172:Complement C1 / Q000096:biosynthesis; D015923:Complement C1r; D006680:HLA Antigens / Q000096:biosynthesis; D006801:Humans; D009504:Neutrophils / Q000378:metabolism* / Q000502:physiology; D010587:Phagocytosis; D011499:Protein Processing, Post-Translational; D012313:RNA / Q000302:isolation & purification; D012333:RNA, Messenger / Q000096:biosynthesis*; D011961:Receptors, Fc / Q000096:biosynthesis
D001798:Blood Proteins; D003172:Complement C1; D006680:HLA Antigens; D012333:RNA, Messenger; D011961:Receptors, Fc; D012313:RNA; D003166:Complement Activating Enzymes; D015923:Complement C1r