Structural insights into DNA sequence recognition by Type ISP restriction-modification enzymes

Manasi Kulkarni, Neha Nirwan, Kara van Aelst, Mark Szczelkun, Kayarat Saikrishnan

Research output: Contribution to journalArticle (Academic Journal)peer-review

11 Citations (Scopus)
358 Downloads (Pure)

Abstract

Engineering restriction enzymes with new sequence specificity has been an unaccomplished challenge, presumably because of the complexity of target recognition. Here we report detailed analyses of target recognition by Type ISP restriction-modification enzymes. We determined the structure of the Type ISP enzyme LlaGI bound to its target and compared it with the previously reported structure of a close homologue that binds to a distinct target, LlaBIII. The comparison revealed that, although the two enzymes use almost a similar set of structural elements for target recognition, the residues that read the bases vary. Change in specificity resulted not only from appropriate substitution of amino acids that contacted the bases but also from new contacts made by positionally distinct residues directly or through a water bridge. Sequence analyses of 552 Type ISP enzymes showed that the structural elements involved in target recognition of LlaGI and LlaBIII were structurally well-conserved but sequentially less-conserved. In addition, the residue positions within these structural elements were under strong evolutionary constraint, highlighting the functional importance of these regions. The comparative study helped decipher a partial consensus code for target recognition by Type ISP enzymes.
Original languageEnglish
Pages (from-to)4396-4408
Number of pages13
JournalNucleic Acids Research
Volume44
Issue number9
Early online date14 Mar 2016
DOIs
Publication statusPublished - 19 Mar 2016

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