Secondary structure of Tetrahymena thermophilia 5S ribosomal RNA as revealed by enzymatic digestion and microdensitometric analysis.
The secondary structure of [32P] end-labeled 5S rRNA from Tetrahymena thermophilia (strain B) has been investigated using the enzymes S1 nuclease, cobra venom ribonuclease and T2 ribonuclease. The results, analyzed by scanning microdensitometry and illustrated by three-dimensional computer graphics, support the secondary structure model of Curtiss and Vournakis for 5S rRNA. Aberrent mobility of certain RNA fragments on sequencing gels was observed as regions of band compression. These regions are postulated to be caused by stable internal base-pairing. The molecule was probed with T2 RNase in neutral (pH 7.5) and acidic (pH 4.5) buffers and only minor structural differences were revealed. One of the helices was found to be susceptible to enzymatic attack by both the single-strand and double-strand specific enzymes. These observations are evidence for the existence of dynamic structural equilibria in 5S rRNA.
D003720:Densitometry; D004720:Endonucleases; D004722:Endoribonucleases; D009690:Nucleic Acid Conformation; D012335:RNA, Ribosomal; D012259:Ribonuclease, Pancreatic; D012270:Ribosomes / Q000648:ultrastructure*; D015719:Single-Strand Specific DNA and RNA Endonucleases; D013768:Tetrahymena / Q000648:ultrastructure*
D012335:RNA, Ribosomal; D004720:Endonucleases; D004722:Endoribonucleases; C022543:ribonuclease T(2); D012259:Ribonuclease, Pancreatic; D015719:Single-Strand Specific DNA and RNA Endonucleases