Protein fragment complementation in M.HhaI DNA methyltransferase.
The 5mC DNA methyltransferase M.HhaI can be split into two individually inactive N- and C-terminal fragments that together can form an active enzyme in vivo capable of efficiently methylating DNA. This active fragment pair was identified by creating libraries of M.HhaI gene fragment pairs and then selecting for the pairs that code for an active 5mC methyltransferase. The site of bisection for successful protein fragment complementation in M.HhaI was in the variable region near the target recognition domain between motif VIII and TRD. This same region is the location of bifurcation in the naturally split 5mC methyltransferase M.AquI, the location for circular permutation in M.BssHII, and the location for previously engineered split versions of M.BspRI.
Choe
Wonchae
W
Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 3400 N. Charles St. Baltimore, MD 21218, USA.
Chandrasegaran
Srinivasan
S
Ostermeier
Marc
M
D000595:Amino Acid Sequence; D001665:Binding Sites; D004247:DNA / Q000737:chemistry*; D019175:DNA Methylation; D015257:DNA-Cytosine Methylases / Q000032:analysis / Q000737:chemistry*; D004789:Enzyme Activation; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry* / Q000235:genetics; D011485:Protein Binding; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D013329:Structure-Activity Relationship
D010446:Peptide Fragments; D004247:DNA; D015257:DNA-Cytosine Methylases