Hemin and cyclic AMP stimulate message-dependent translation in lysates from Friend erythroleukemia cells.
A message-dependent, cell-free translation system was prepared from both uninduced and N,N'-hexamethylene-bis-acetamide-induced Friend erythroleukemia cells following modification of standard protocols. Active extracts were prepared by lysing Friend erythroleukemia cells in hypotonic buffer containing 50 microM hemin and 10 mM cyclic adenosine monophosphate (cAMP), and assaying translation in vitro in the absence of exogenous nucleoside triphosphates. Both hemin and cAMP were required for full activity and appeared to act by prolonging initiation in vitro. Following treatment of cell extracts with micrococcal nuclease, brome mosaic virus, globin, equine herpesvirus type 1, and frog virus 3 mRNA were efficiently translated to yield full-sized products. Using brome mosaic virus as a test message, levels of translation equivalent to 30%-50% of that seen in mock-treated lysates were obtained. These results attest to the translational efficiency of extracts prepared and assayed in the manner described above, and suggest that such extracts may be useful both for routine translational studies, and as a tool to dissect translational changes accompanying erythroid differentiation in vitro.
Chinchar
V G
VG
Department of Microbiology, University of Mississippi Medical Center, Jackson 39216.
Turner
L A
LA
Kumar
G
G
D000081:Acetamides / Q000494:pharmacology; D000818:Animals; D002461:Cell Line, Transformed; D002474:Cell-Free System; D000242:Cyclic AMP / Q000494:pharmacology*; D005622:Friend murine leukemia virus; D006418:Heme / Q000031:analogs & derivatives*; D006427:Hemin / Q000494:pharmacology*; D004915:Leukemia, Erythroblastic, Acute / Q000235:genetics*; D014176:Protein Biosynthesis / Q000187:drug effects*
D000081:Acetamides; D006418:Heme; D006427:Hemin; D000242:Cyclic AMP; C014026:hexamethylene bisacetamide