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Sequence requirements for splicing by the Cne PRP8 intein
Author(s) -
Pearl Esther J.,
Tyndall Joel D.A.,
Poulter Russell T.M.,
Wilbanks Sigurd M.
Publication year - 2007
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.2007.05.060
Subject(s) - intein , protein splicing , rna splicing , splice , cleavage (geology) , genetics , mutant , context (archaeology) , histidine , alanine scanning , biology , computational biology , biochemistry , chemistry , gene , mutagenesis , amino acid , rna , paleontology , fracture (geology)
The dependence of protein splicing on conserved residues of the Cne PRP8 intein was assessed by alanine scanning mutagenesis in a foreign protein context. Corroboration was obtained for the involvement of residues at the splice junctions and of the conserved threonine and histidine of motif B. Five additional residues were identified as absolutely required for splicing. Variant W151A displayed premature C‐terminal cleavage, not seen with other Cne PRP8 mutants. We propose a model whereby W151 acts to prevent premature C‐terminal cleavage, favoring complete splicing as opposed to two disjointed cleavage events.