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A2 Transfer RNA ( tRNA ). Role in translation Structure Synthesis and processing Modifications and nucleotides. 1. Role in translation. Transfer RNAs (tRNAs) are small molecules that act as adaptors during protein synthesis.
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Role in translation • Structure • Synthesis and processing • Modifications and nucleotides
1. Role in translation • Transfer RNAs (tRNAs) are small molecules that act as adaptors during protein synthesis. • nucleotide sequence of mRNA---tRNA---amino acid sequence of the polypeptide • Each tRNA bind a specific amino acid, recognize its codon in mRNA, place this amino acid in the growing polypeptide.
3’ 3’ ACC ACC 5’ 5’
2. tRNA structure • Primary structure: 74-95nt, with modified nucleotides • Secondary structure: cloverleaf['kləuvəli:f] composed of a series of stem-loop structure known as arms. • acceptor arm (氨基酸臂): CCA as the attachment site for amino acid • D or DHU arm (二氢尿嘧啶环): dihydrouracil • anticodon arm (反密码子环): recognize and bind codon in the mRNA
optional arm or extra or variable arm: Class I tRNA, 2-3 nucleotides; Class ⅡtRNA, 13-21 nucleotides, up to five base pairs • TψC arm: contains Tψ • Invariant, semi-variant and not conserved nucleotides • 3D-structure(tertiary structure): acceptor arm and anticodon loop are at opposite ends.
3. Synthesis and processing • Synthesis: RNA polymerase Ⅲ transcribe tRNA genes which exist as multiple copies • Processing: RNase P and RNase D • Several tRNA genes may be transcribed together as a single pre-tRNA which is then processed by ribonucleases that cleave at the 5’ and 3’ ends of each tRNA sequence.
In prokaryotes, processing is carried out by a series order of steps by ribonucleases (RNase D and RNase P) and RNA component of RNase P has catalytic activity known as ribozyme. • CCA (the attachment site for amino acid) is present in the coding sequence although sometimes is removed by RNase D.
In eukaryotes, introns in pre-tRNA transcript must be removed during processing and CCA is not present in eukaryotic tRNA gene and is added later.
4.Modification of nucleotides: tRNA contains modified bases that influence its pairing properties • Methylation of the ribose sugar of the nucleotide. e.g: 7-methylguanosine. • Base rearrangements: atom position interchanging. e.g: uradine to pseudouradine • Double-bond saturation: uradine to dihydrouridine • Deamination: guanosine to inosine • Sulfur substitution: uradine to 4-thiouradine (硫尿核苷) • Addition of larger groups: guanosine to queosine
Key points • The role of tRNA in translation • Processing and modifications of tRNA