Specificity of Mnt 'master residue' obtained from in vivo and in vitro selections

Fauzi S. Silbaq, Steven E. Ruttenberg, Gary D. Stormo

Research output: Contribution to journalReview articlepeer-review

6 Scopus citations

Abstract

Mnt is a repressor from phage P22 that belongs to the ribbon-helix-helix family of DNA binding factors. Four amino acids from the N-terminus of the protein, Arg2, His6, Asn8 and Arg10, interact with the base pairs of the DNA to provide the sequence specificity. Raumann et al. (Nature Struct. Biol., 2, 1115-1122) identified position 6 as a 'master residue' that controls the specificity of the protein. Models for the interaction have residue 6 of Mnt interacting directly with position 5 of the operator. In vivo selections demonstrated that protein variants at residue 6 bound specifically to operator mutations at that position. Operators in which the wild-type G at position 5 was replaced by T specifically bound to several different protein variants, primarily hydrophobic residues. The obtained protein variants, plus some others, were used in in vitro selections to determine their preferred binding sites. The results showed that the residue at position 6 influenced the preference for binding site bases predominantly at position 5, but that the effects of altering it can extend over longer distances, consistent with its designation as a 'master residue'. The similarities of binding sites for different residues do not correlate strongly with common measures of amino acid similarities.

Original languageEnglish
Pages (from-to)5539-5548
Number of pages10
JournalNucleic acids research
Volume30
Issue number24
DOIs
StatePublished - Dec 15 2002

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