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Comparative single‐turnover kinetic analyses of trans ‐cleaving hammerhead ribozymes with naturally derived non‐conserved sequence motifs
Author(s) -
Weinberg Marc S.,
Rossi John J.
Publication year - 2005
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2005.02.016
Subject(s) - ribozyme , hammerhead ribozyme , hairpin ribozyme , cleavage (geology) , mammalian cpeb3 ribozyme , ligase ribozyme , vs ribozyme , biology , chemistry , stereochemistry , rna , genetics , gene , paleontology , fracture (geology)
trans ‐Cleaving hammerhead ribozyme variants were generated with mimicked non‐conserved internal loop motifs derived from five structurally diverse natural cis ‐cleaving ribozymes. Most modified trans ‐cleaving variants showed enhanced single‐turnover cleavage rates relative to minimal counterparts that lack tertiary interactions between internal loop motifs I and II, and relative to controls with sequence changes in loop I. The trans ‐cleaving ribozyme derived from the positive strand of peach latent mosaic viroid had the highest observed cleavage rate, suggesting a structurally optimized motif that facilitates rapid formation of the ribozyme catalytic center in a trans ‐reaction.