title: Unusual ribosome binding properties of mRNA encoding bacteriophage lambda repressor.
abstract: The mRNA encoding repressor cI of phage lambda is the only known E. coli message which starts directly with the initiation AUG codon. The ability of in vitro synthesized cI mRNA fragments (150 or 400 nts) to form ternary initiation complexes has been studied using the toeprint method. In the presence of tRNA(Met)f, these fragments are capable of forming the ternary complexes at the 5'-terminal AUG codon not only with 30S subunits but also with undissociated 70S ribosomes (70S tight couples). In the latter case, no binding at other positions of cI mRNA can be detected at all. The starting region of cI mRNA has a single stranded conformation and is highly enriched in A-residues. This feature of cI mRNA RBS is suggested to be the main factor which allows cI mRNA to form the initiation complex with the ribosome. Unlike 30S subunits, the binding to 70S tight couples is not affected by any of the initiation factors, although it is as efficient as that to 30S subunits supplemented with the factors. 30S subunits prefer to associate with the internal RBSs of the preformed mRNA molecules, provided that they are not sequestered by the secondary structure. In contrast, 70S tight couples tend to avoid extra sequences upstream of the codon directed to the P site and occupy a position as close as possible to the 5'-end of the message. This has been found to be the case both for tRNA(Met)f and for elongator tRNA(Glu)2. The structural features of mRNA RBSs which influence their different binding for 30S subunits and 70S ribosomes are discussed.
authors.affiliation: A.N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Russia.
authors.forename: A G
authors.initials: AG
authors.lastname: Balakin
authors.forename: E A
authors.initials: EA
authors.lastname: Skripkin
authors.forename: I N
authors.initials: IN
authors.lastname: Shatsky
authors.forename: A A
authors.initials: AA
authors.lastname: Bogdanov
mesh_terms: D010582:Bacteriophage lambda / Q000235:genetics; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins; D004926:Escherichia coli / Q000235:genetics; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D012333:RNA, Messenger / Q000378:metabolism*; D012343:RNA, Transfer / Q000378:metabolism; D012367:RNA, Viral / Q000378:metabolism*; D012097:Repressor Proteins / Q000235:genetics*; D012270:Ribosomes / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics*; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins
chemical_list: D004268:DNA-Binding Proteins; D012333:RNA, Messenger; D012367:RNA, Viral; D012097:Repressor Proteins; D014157:Transcription Factors; D014764:Viral Proteins; D054334:Viral Regulatory and Accessory Proteins; C016649:phage repressor proteins; D012343:RNA, Transfer