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Protein Splicing of inteins from Synechococcus sp. PCC 7002 and Pyrococcus abyssi
Author(s) -
Karanja Kevin K.,
Urbanski Laura M.,
Reitter Julie N.,
Mills Kenneth V.
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.756.5
Subject(s) - intein , protein splicing , rna splicing , biology , mutant , biochemistry , genetics , chemistry , gene , rna
Protein splicing is a four‐step intramolecular reaction, in which an intein flanked by N‐ and C‐terminal exteins is removed, with ligation of the exteins. We are studying inteins that interrupt genes from Synechococcus sp. PCC 7002 ( Ssp 7002 ), Trichodesmium erythraeum ( Tery ) or Pyrococcus abyssi DNA Polymerase II ( Pab Pol II). The cyanobacterium Ssp 7002 has an intein that interrupts a putative dTDP glucose dehydratase. The Ssp 7002 intein has a highly conserved C‐terminal Asn, unlike an intein from Tery , which has 63% sequence identity with the Ssp 7002 intein but has a C‐terminal Gln. Both inteins splice with their native C‐terminal residue. We plan to study the effect of switching Gln for Asn (and vice versa), as well as the influence of conserved residues on the mechanism of splicing. The Pyrococcus abysii PolII intein has a conserved, catalytic His residue. Mutation of this residue to Ala or Gly prevents splicing. We will explore whether we can rescue activity in the His‐Gly mutant in vivo by incubation with imidazole derivatives. This material is based upon work supported by the National Science Foundation under grant MCB‐0950245 and by the Camille and Henry Dreyfus Foundation.