[Adequate system for investigation of translation initiation of the human retrotransposon L1 mRNA in vitro].
Retrotransposon L1 codes for a unique dicistronic mRNA which serves both a transposition intermediate and a template for the synthesis of two proteins of this mobile element. According to preliminary data, the translation initiation of both cistrons of L1 occurs by non-canonical mechanisms. When translating the L1 mRNA in rabbit reticulocyte lysate (RRL), a standard system routinely used by many researchers to study mechanisms of translation initiation in eukaryotes, we observed along with expected products a number of polypeptides resulted from aberrant initiation at internal AUG codons. Such products are absent on translation of L1 mRNA in vivo. Addition to the system of a cytoplasmic extract from HeLa cells resulted in disappearance of these abberant products whereas the efficiency of translation of the first cistron remained unchanged. The level of translation of the second cistron became significantly lower. This also made the picture closer to that observed in vivo. These and other experiments allowed us to clearly demonstrate that the new combined cell-free system is much more adequate to study mechanisms of translation initiation than a regular RRL.
Dmitriev
S E
SE
Bykova
N V
NV
Andreev
D E
DE
Terenin
I M
IM
D000818:Animals; D002474:Cell-Free System; D018387:Codon, Initiator; D006367:HeLa Cells; D006801:Humans; D066298:In Vitro Techniques; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics* / Q000502:physiology; D011817:Rabbits; D012156:Reticulocytes / Q000378:metabolism; D018626:Retroelements / Q000502:physiology
D018387:Codon, Initiator; D012333:RNA, Messenger; D018626:Retroelements