程序性核糖体移码案例.ppt

* The research of Programmed –1 ribosomal frameshifting (-1PRF) lecturer ： INTRODUCTION Programmed ribosomal frameshifting (PRF) is a recoding event that leads to a change of the translational reading frame and allows the translation of messenger RNAs (mRNAs)with overlapping open reading frames, often yielding two protein products from one mRNA. During PRF, the reading frame may shift in +1, –1, or even –2 direction. The mechanisms of +1 and –1PRF appear to be quite different and require a different set of stimulatory elements. INTRODUCTION The classic example of -1PRF is slippage triggered by two elements in the mRNA, a ‘‘slippery sequence’’ and a downstream secondary structure element. The slippery site is made up by sequences such as X XXY YYZ, where XXY and YYZ are codons in the original frame (0 frame) that presumably favor tRNA sliding on the mRNA and anticodon misalignment, changing the codons to XXX and YYY in the –1 frame. INTRODUCTION The stimulatory secondary structure elements of the mRNA, which usually consist of a stem loop or a pseudoknot located at a particular distance downstream of the slippery sequence, may enhance –1PRF by inducing pausing of the ribosome or by perturbing normal decoding. The mechanism of –1PRF is not clear, and there are at least four groups of models as to when and why it occurs. RESULTS Pathway I, –1PRF occurs during translocation (TL) of tRNA nnX (gray) and tRNA XXY (green). Pathway II, –1PRF after correct reading the 0 frame codon and upon accommodation (Acc) of tRNA YYZ (magenta); the following codon is read in –1 frame. Pathway III, during translocation of tRNA XXY and tRNA YYZ that exposes –1 frame codon Zab in the A site. Pathway IV, upon decoding of the codon following the slippery sequence; 0 frame tRNA is shown in blue RESULTS 1. Efficiency of –1PRF In Vivo and In Vitro Time courses of synthesis of fMetTyr (MY), fMetTyrLeu (MYL), fMetTyrLeuLys (MYLK), fMetTyrLeuLysPhe (MYLKF), and fMetTyrLeuLysVal (MYLKV) u