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Cell & Molecular
Biology
Ribozymes
Chapter 7
RNA with catalytic activity
Self-Splicing RNA
Self-splicing intron
        of bacterial tRNA



http://www.pdb.org/pdb/101/motm.do?momID=65
Self-Cleaving RNA
A riboswitch that
      becomes a self-
      cleaving ribozyme
      in the presence of
      glucosamine.


http://www.pdb.org/pdb/101/motm.do?momID=130
Riboswitch
“...regulatory elements
built directly into
a messenger RNA.”



         http://www.pdb.org/pdb/101/motm.do?momID=130
Guanine riboswitch



http://www.pdb.org/pdb/101/motm.do?momID=130
RESEARCH ARTICLE
                                                                                                   sequence segments (Fig. 1B). The
             RNA-Catalyzed RNA                                                                     nus of the ligase domain was cov
                                                                                                   tached to an RNA primer so that

         Polymerization: Accurate and                                                              able to catalyze primer extension
                                                                                                   selected by virtue of their attachm
                                                                                                   primer that they extended.
        General RNA-Templated Primer                                                                   In contrast to the parental
                                                                                                   which hybridizes to the template, a
                  Extension                                                                        capable of general polymerization
                                                                                                   ognize the primer-template compl
           Wendy K. Johnston, Peter J. Unrau,* Michael S. Lawrence,                                relying on sequence-specific in
                    Margaret E. Glasner, David P. Bartel†                                          Therefore, the template RNA was d
                                                                                                   be too short for extensive hybridiz
                                                                                                   the ribozyme (Fig. 1B). For the p
      The RNA world hypothesis regarding the early evolution of life relies on the                 bozyme (Fig. 1A), the pairing be
      premise that some RNA sequences can catalyze RNA replication. In support of                  ribozyme and the template also co
      this conjecture, we describe here an RNA molecule that catalyzes the type of                 stem known to be necessary for li
      polymerization needed for RNA replication. The ribozyme uses nucleoside                      tion (16). To restore this stem, a s
      triphosphates and the coding information of an RNA template to extend an RNA                 GGCACCA ( purple RNA in Fig
      primer by the successive addition of up to 14 nucleotides—more than a com-                   introduced to hybridize to the segm
      plete turn of an RNA helix. Its polymerization activity is general in terms of the           ligase domain that formerly paire
      sequence and the length of the primer and template RNAs, provided that the                   template. Finally, because a mo
      3 terminus of the primer pairs with the template. Its polymerization is also                 mode of primer-template recogni
      quite accurate: when primers extended by 11 nucleotides were cloned and                      require the participation of an
      sequenced, 1088 of 1100 sequenced nucleotides matched the template.                          RNA domain, a 76-nt random-seq
                                                                                                   ment was appended to the 3 term
The RNA world hypothesis states that early life   join oligonucleotides assembled on a tem-        degenerate ligase domain (Fig. 1B
forms lacked protein enzymes and depended         plate (10–12). However, the templates that           Sequence variants able to rec
instead on enzymes composed of RNA (1).           can be copied are limited to those that match    primer-template in this new configu
Much of the appeal of this hypothesis comes       the oligonucleotide substrates, and it has not   then extend the primer with tagged
from the realization that ribozymes would have    been possible to include sufficient concentra-   were enriched by repeated rounds
been far easier to duplicate than proteinaceous   tions of all the oligonucleotide substrates      selection and amplification (Table
enzymes (2–5). Whereas coded protein replica-     needed for a general copying reaction. More      that extended their primer by using
Artificial
Ribozymes
Table 7-4 Essential Cell Biology (© Garland Science 2010)
The RNA World
Figure 7-43 Essential Cell Biology (© Garland Science 2010)
Figure 7-44 (part 1 of 3) Essential Cell Biology (© Garland Science 2010)
Figure 7-44 (part 2 of 3) Essential Cell Biology (© Garland Science 2010)
Figure 7-44 (part 3 of 3) Essential Cell Biology (© Garland Science 2010)
Figure 7-45 Essential Cell Biology (© Garland Science 2010)
Figure 7-42 Essential Cell Biology (© Garland Science 2010)
Figure 7-46 Essential Cell Biology (© Garland Science 2010)

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Ribozymes

  • 2.
  • 3.
  • 6.
  • 7.
  • 10. Self-splicing intron of bacterial tRNA http://www.pdb.org/pdb/101/motm.do?momID=65
  • 12. A riboswitch that becomes a self- cleaving ribozyme in the presence of glucosamine. http://www.pdb.org/pdb/101/motm.do?momID=130
  • 14. “...regulatory elements built directly into a messenger RNA.” http://www.pdb.org/pdb/101/motm.do?momID=130
  • 16. RESEARCH ARTICLE sequence segments (Fig. 1B). The RNA-Catalyzed RNA nus of the ligase domain was cov tached to an RNA primer so that Polymerization: Accurate and able to catalyze primer extension selected by virtue of their attachm primer that they extended. General RNA-Templated Primer In contrast to the parental which hybridizes to the template, a Extension capable of general polymerization ognize the primer-template compl Wendy K. Johnston, Peter J. Unrau,* Michael S. Lawrence, relying on sequence-specific in Margaret E. Glasner, David P. Bartel† Therefore, the template RNA was d be too short for extensive hybridiz the ribozyme (Fig. 1B). For the p The RNA world hypothesis regarding the early evolution of life relies on the bozyme (Fig. 1A), the pairing be premise that some RNA sequences can catalyze RNA replication. In support of ribozyme and the template also co this conjecture, we describe here an RNA molecule that catalyzes the type of stem known to be necessary for li polymerization needed for RNA replication. The ribozyme uses nucleoside tion (16). To restore this stem, a s triphosphates and the coding information of an RNA template to extend an RNA GGCACCA ( purple RNA in Fig primer by the successive addition of up to 14 nucleotides—more than a com- introduced to hybridize to the segm plete turn of an RNA helix. Its polymerization activity is general in terms of the ligase domain that formerly paire sequence and the length of the primer and template RNAs, provided that the template. Finally, because a mo 3 terminus of the primer pairs with the template. Its polymerization is also mode of primer-template recogni quite accurate: when primers extended by 11 nucleotides were cloned and require the participation of an sequenced, 1088 of 1100 sequenced nucleotides matched the template. RNA domain, a 76-nt random-seq ment was appended to the 3 term The RNA world hypothesis states that early life join oligonucleotides assembled on a tem- degenerate ligase domain (Fig. 1B forms lacked protein enzymes and depended plate (10–12). However, the templates that Sequence variants able to rec instead on enzymes composed of RNA (1). can be copied are limited to those that match primer-template in this new configu Much of the appeal of this hypothesis comes the oligonucleotide substrates, and it has not then extend the primer with tagged from the realization that ribozymes would have been possible to include sufficient concentra- were enriched by repeated rounds been far easier to duplicate than proteinaceous tions of all the oligonucleotide substrates selection and amplification (Table enzymes (2–5). Whereas coded protein replica- needed for a general copying reaction. More that extended their primer by using
  • 17.
  • 18.
  • 20. Table 7-4 Essential Cell Biology (© Garland Science 2010)
  • 22. Figure 7-43 Essential Cell Biology (© Garland Science 2010)
  • 23. Figure 7-44 (part 1 of 3) Essential Cell Biology (© Garland Science 2010)
  • 24. Figure 7-44 (part 2 of 3) Essential Cell Biology (© Garland Science 2010)
  • 25. Figure 7-44 (part 3 of 3) Essential Cell Biology (© Garland Science 2010)
  • 26. Figure 7-45 Essential Cell Biology (© Garland Science 2010)
  • 27. Figure 7-42 Essential Cell Biology (© Garland Science 2010)
  • 28. Figure 7-46 Essential Cell Biology (© Garland Science 2010)