A Neurospora crassa heat-shocked cell lysate translates homologous and heterologous messenger RNA efficiently, without preference for heat shock messages.
Cell-free protein synthesis systems were prepared from normally-grown (N-lysate) and heat-shocked (HS-lysate) Neurospora crassa mycelium. Although both lysates translated homologous mRNA, the HS-lysate was more active, yielding a higher incorporation of [35S]-methionine into hot TCA-insoluble material and a vastly superior protein synthesis profile. The optimal temperature for translation by both lysates was 21 degrees C; the HS-lysate did not translate heat-shock mRNA preferentially at any temperature tested. Fortuitously, heterologous messenger RNAs from diverse eukaryotic and viral sources - Drosophila, dog pancreas, rabbit globin mRNA, brome mosaic virus, tobacco mosaic virus - were translated by the HS-lysate with an efficiency comparable to that of the commercial rabbit reticulocyte system and superior to the wheat germ system. The cap analogues, m7G(5')ppp(5')G and m7G(5')Gm, inhibited translation significantly.
Curle
C A
CA
Department of Biological Sciences, University of Calgary, Alberta, Canada.
Kapoor
M
M
D000818:Animals; D002474:Cell-Free System; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification; D006358:Hot Temperature; D007700:Kinetics; D009491:Neurospora / Q000235:genetics*; D009492:Neurospora crassa / Q000235:genetics*; D014176:Protein Biosynthesis; D012315:RNA Caps / Q000235:genetics; D012331:RNA, Fungal / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics*; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism
D005656:Fungal Proteins; D012315:RNA Caps; D012331:RNA, Fungal; D012333:RNA, Messenger