Selected article for: "cleavage site and vitro assay"
Author: Aaron S. Mendez; Carolin Vogt; Jens Bohne; Britt A. Glaunsinger
Title: Site specific target binding controls RNA cleavage efficiency by the Kaposi’s sarcoma-associated herpesvirus endonuclease SOX
  • Document date: 2018_5_13
    • ID: 298cbr1x_19
    Snippet: Finally, we sought to verify that the predominant 27 nt cleavage product we observed was a result of an endonucleolytic cleavage and not 5' end processing. To this end, we generated a LIMD1 54 substrate containing a 3' 32 P pCp label and a free 5' OH to block 5' end processing. Again, in the presence of SOX, WT LIMD1 54 but not the A-G mutant produced a cleavage product whose size corresponded to cleavage at the predicted site ( Fig 6B) . Taken t.....
    Document: Finally, we sought to verify that the predominant 27 nt cleavage product we observed was a result of an endonucleolytic cleavage and not 5' end processing. To this end, we generated a LIMD1 54 substrate containing a 3' 32 P pCp label and a free 5' OH to block 5' end processing. Again, in the presence of SOX, WT LIMD1 54 but not the A-G mutant produced a cleavage product whose size corresponded to cleavage at the predicted site ( Fig 6B) . Taken together, these data confirm that our in vitro assay faithfully recapitulates SOX cleavage site specificity on a true substrate. The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/320929 doi: bioRxiv preprint

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