A poly(A) tail-responsive in vitro system for cap- or IRES-driven translation from HeLa cells.
In cells, the poly(A) tail stimulates translation from messenger RNAs bearing a cap structure or viral IRES elements. This 3' end-mediated stimulation of translation is not reflected in commonly used commercial cell-free translation systems prepared from rabbit reticulocytes or wheat germ. We describe a simple procedure to generate poly(A) tail-responsive translation extracts from HeLa cells. We suggest that this procedure should be adaptable to many animal cell lines.
Thoma
Christian
C
Gene Expression Programme, EMBL, Heidelberg, Germany.
Ostareck-Lederer
Antje
A
Hentze
Matthias W
MW
D002474:Cell-Free System; D004686:Encephalomyocarditis virus / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D010442:Peptide Chain Initiation, Translational; D011061:Poly A / Q000737:chemistry / Q000378:metabolism*; D014176:Protein Biosynthesis; D012315:RNA Caps; D012333:RNA, Messenger / Q000737:chemistry / Q000235:genetics*; D012367:RNA, Viral / Q000737:chemistry / Q000235:genetics*; D012270:Ribosomes / Q000502:physiology
D012315:RNA Caps; D012333:RNA, Messenger; D012367:RNA, Viral; D011061:Poly A